CN108948208A - 一种可注射自修复水下蛋白及其用途 - Google Patents
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Abstract
本发明涉及水下粘合材料领域,特别是涉及一种可注射自修复水下蛋白及其用途。本发明提供一种融合蛋白,所述融合蛋白包括贻贝足丝蛋白片段和卷曲螺旋结构片段。本发明将贻贝足丝蛋白与卷曲螺旋结构进行基因重组,从而提供了一种新的融合蛋白,所述融合蛋白可以在溶液中自组装形成凝胶材料,兼具贻贝足丝蛋白的界面粘性以及卷曲螺旋结构的内在粘性,从而实现水下超粘合强度。
Description
技术领域
本发明涉及水下粘合材料领域,特别是涉及一种可注射自修复水下蛋白及其用途。
背景技术
水下粘合材料作为一种新型的功能性材料,近年来逐步受到广泛关注,被日益增多的领域所需求。其中最典型的是生物医药、海洋工程领域。在生物医药领域,传统的治疗手段会使用手术螺钉以及缝线方式来进行伤口缝合。然而这些手段通常会使患者承受相当大的痛苦,且治愈后必须进行拆除,往往会带来炎症和感染的隐患。因而研究人员把目光投向粘合材料,希望在此基础上创造出新型伤口治疗的手段。而由于人体组织处于一种非常复杂的水环境中,因而开发出在较高湿度或水环境中依然能保持粘合特性的医用材料有着重要意义。在海洋工程领域,船舶的修补,海底输油管道及江河大坝裂缝的填充都离不开水下粘合材料的发展,这使得人们对性能优越的水下粘合材料的需求愈发迫切。
尽管水下粘合材料为社会所需,制备多功能高粘合强度的水下粘合材料仍是困扰人们的难题。然而,海洋中的一些生物给研究人员带来诸多灵感。通过对海洋或水下生物如贻贝、沙塔蠕虫、石蛾、藤壶等生物所分泌的粘性蛋白进行研究,许多粘性蛋白的水下粘合机制被逐渐揭示,越来越多的仿生水下粘合材料应运而生。这方面的研究包括:1)基于邻苯二酚结构的仿生水下粘合材料研究:利用化学方法将邻苯二酚结构基团修饰于高分子的主链中;2)基于凝聚复合结构的仿生水下粘合材料研究:利用带相反电荷的分子,将其混合后,在静电作用、疏水作用下所形成的球形颗粒团聚物,起到水下粘合的作用;3)基于淀粉样蛋白结构的仿生水下粘合材料研究:利用基因工程技术将贻贝足丝蛋白(Mfp-3和Mfp-5)同大肠杆菌淀粉样蛋白卷曲纤维亚基成分CsgA融合,构成重组蛋白CsgA-Mfp3和Mfp5-CsgA,使两种蛋白模块同时发挥出粘合特性。虽然天然的水下粘合材料主要由蛋白质组成,然而现阶段多数仿生水下粘合材料大部分是基于化学合成方法制备而成,在这些分子中注入的水下粘合特征较为单一,因而制备的粘合材料在水环境下并不具有自然粘合材料的高粘合强度。此外,较差的可注射性也限制了这些材料在各种领域特别是生物医药领域中的广泛应用。
发明内容
鉴于以上所述现有技术的缺点,本发明的目的在于提供一种融合蛋白及含有所述融合蛋白的胶体的制备方法和用途,用于解决现有技术中的问题。
为实现上述目的及其他相关目的,本发明第一方面提供一种融合蛋白,所述融合蛋白包括贻贝足丝蛋白片段和卷曲螺旋结构片段。
在本发明一些实施方式中,所述贻贝足丝蛋白选自紫贻贝足丝蛋白或地中海贻贝足丝蛋白。
在本发明一些实施方式中,所述贻贝足丝蛋白选自mefp-3、mefp-5、Mgfp-3、Mgfp-5、mfp-3s或mfp3s-pep。
在本发明一些实施方式中,所述贻贝足丝蛋白片段为:a)氨基酸序列如SEQ IDNo.4-9所示的多肽片段;
或b)氨基酸序列与SEQ ID NO.4-9具有80%以上同源性、且具有a)限定的多肽片段的功能的多肽片段。
在本发明一些实施方式中,所述贻贝足丝蛋白来源于贻贝。
在本发明一些实施方式中,所述卷曲螺旋结构片段选自亮氨酸拉链片段、A1片段、ZE/ZR片段、GCN4片段、γ52-88KI片段、[(AG)3PEG]10片段、PCxP片段、NLP片段。
在本发明一些实施方式中,所述亮氨酸拉链片段包括A片段、S片段和P片段;
所述A片段为:c)氨基酸序列如SEQ ID No.1所示的多肽片段;或d)氨基酸序列与SEQ ID NO.1具有80%以上同源性、且具有c)限定的多肽片段的功能的多肽片段;
所述S片段为:e)氨基酸序列如SEQ ID No.2所示的多肽片段;或f)氨基酸序列与SEQ ID NO.2具有80%以上同源性、且具有e)限定的多肽片段的功能的多肽片段;
所述P片段为:g)氨基酸序列如SEQ ID No.3所示的多肽片段;或h)氨基酸序列与SEQ ID NO.3具有80%以上同源性、且具有g)限定的多肽片段的功能的多肽片段。
在本发明一些实施方式中,所述融合蛋白还包括功能性蛋白片段。
在本发明一些实施方式中,所述亮氨酸拉链片段自N端至C端依次包括A片段、S片段和P片段。
在本发明一些实施方式中,所述贻贝足丝蛋白片段被插入亮氨酸拉链片段中。
在本发明一些实施方式中,所述融合蛋白自N端至C端依次包括A片段、S片段、贻贝足丝蛋白片段和P片段。
在本发明一些实施方式中,所述融合蛋白经酪氨酸酶修饰。
在本发明一些实施方式中,所述功能性蛋白片段选自粘性蛋白片段、细胞粘附多肽片段、酶蛋白片段、生长因子片段、抗菌活性多肽片段、凝血因子片段、生物相容性多肽片段、抗生肽片段、监测多肽片段中的一种或多种的组合。
在本发明一些实施方式中,所述融合蛋白自N端至C端依次包括A片段、S片段、功能性蛋白片段、贻贝足丝蛋白片段和P片段。
本发明第二方面提供一种分离的多核苷酸,编码如上所述的融合蛋白。
本发明第三方面提供一种构建体,所述构建体含有如上所述的分离的多核苷酸。
本发明第四方面提供一种表达系统,所述表达系统含有如上所述的构建体或基因组中整合有外源的如上所述的多核苷酸。
本发明第五方面提供所述的融合蛋白的制备方法,包括:在适合表达所述融合蛋白的条件下,培养如上所述的表达系统。
在本发明一些实施方式中,将表达所得的融合蛋白进行酪氨酸酶修饰。
本发明第六方面提供一种自组装胶体溶液,所述溶液包括如上所述的融合蛋白。
本发明第七方面提供一种胶体,所述胶体包括如上所述的融合蛋白。
在本发明一些实施方式中,所述胶体为水凝胶。
在本发明一些实施方式中,胶体中融合蛋白的含量为70mg/mL~250mg/mL。
在本发明一些实施方式中,所述胶体由所述融合蛋白自组装获得。
在本发明一些实施方式中,所述胶体由权利要求9所述的自组装胶体溶液制备获得。
在本发明一些实施方式中,所述胶体中,融合蛋白中的贻贝足丝蛋白选自mefp-3和mefp-5的组合。
本发明第八方面提供所述的融合蛋白、所述的自组装胶体溶液、所述的胶体在水下粘合剂制备领域的用途。
附图说明
图1为水下粘合材料基因模块化构建方法示意图。
图2为构建水下粘合材料粘合原理示意图。
图3为构建的重组A-S-P质粒图谱。
图4为构建的重组A-S-Mefp3-P质粒图谱。
图5为构建的重组A-S-Mefp5-P质粒图谱。
图6为构建的重组A-S-Spytag-Mefp3-P质粒图谱。
图7为构建的重组A-S-Snooptag-Mefp5-P质粒图谱。
图8为蛋白考马斯亮蓝染色实验及免疫印迹鉴定实验验证A-S-Mefp3-P、A-S-Mefp5-P、A-S-P的成功表达及提纯。其中(a)A-S-Mefp3-P、A-S-Mefp5-P考马斯亮蓝染色(左)及免疫印迹鉴定(右)(b)标准蛋白标记物(c)A-S-P考马斯亮蓝染色(左)及免疫印迹鉴定(右)。
图9(a)为圆二色谱实验中圆二色谱谱图。
图9(b)为圆二色谱实验中二级结构所占百分比。
图10为氯化硝基四氮唑蓝染色验证修饰后蛋白的多巴结构。图AS3P、AS5P分别代表未经酪氨酸酶修饰的A-S-Mefp3-P和A-S-Mefp5-P蛋白样品,而AS3P MO、AS5P MO分别代表经酪氨酸酶修饰过的A-S-Mefp3-P和A-S-Mefp5-P蛋白样品。
图11为AFM表征A-S-P系列蛋白的形貌。其中(a)、(b)分别为A-S-Mefp3-P蛋白未经酪氨酸酶修饰和经修饰的形貌,(c)、(d)分别为A-S-Mefp5-P蛋白未经酪氨酸酶修饰和经修饰的形貌,(e)、(f)分别为等质量比A-S-Mefp3-P和A-S-Mefp5-P混合蛋白未经酪氨酸酶修饰和经修饰的形貌,(g)为A-S-P蛋白形貌。
图12为A-S-P系列蛋白水凝胶图。图中ASP指A-S-P蛋白所成水凝胶,A3、A3 MO分别指未经酪氨酸酶修饰和经修饰的A-S-Mefp3-P蛋白所成水凝胶,A5、A5 MO分别指未经酪氨酸酶修饰和经修饰的A-S-Mefp5-P蛋白所成水凝胶,A35和A35 MO分别指未经酪氨酸酶修饰和经修饰的A-S-Mefp3-P和A-S-Mefp5-P蛋白等质量混合所成水凝胶。
图13为ASP系列粘合力搭接剪切测试(a)为ASP系列粘合强度(b)为搭接剪切测试环境。
图14为A35 MO和ASP水下粘合力搭接剪切测试(a)为ASP系列粘合强度(b)为搭接剪切测试环境。
图15为骨粘合实验测试。
图16为A35 MO10与A35 MO粘合力测试对照图。其中黄色柱体代表测试环境为空气湿度30%,紫色柱体代表测试环境为水环境。
图17为ASP和A35 MO冷冻干燥后的SEM图像。其中(a)和(b)为ASP的剖面SEM图,(c)和(d)为A35 MO的剖面SEM图。(a)和(c)标尺为20μm,(b)和(d)标尺为10μm。
图18为ASP和A35 MO振幅扫描测试。
图19为ASP和A35 MO频率扫描测试。其中红色代表A35 MO,黑色代表ASP;实心点为G’,空心点为G”。
图20为A-S-Mefp3-P和A-S-Mefp5-P相互作用的探究实验。(a)为ASpy,(b)为ASnoop,(c)为将Aspy Asnoop相互靠近(d)为荧光显微镜下界面状态(e)为10分钟后界面状态。
图21为A35 MO自修复性质探究实验。(a)为完整的A35 MO(b)为用刀片从水凝胶中间位置切开为两半后的形态(c)为将两部分水凝胶分离后的形态(d)为将两部分水凝胶贴近后融合的形态(e)为将融合后的水凝胶挑起的状态。
图22为A35 MO自修复能力在聚氨酯多孔材料的修复中的运用。
图23为可注射的A35 MO展示(a)及3D打印图案(b)。
图24为利用A35 MO可注射性质粘合玻璃小球填补缺失的材料。
图25为A35 MO水下粘合具体应用实例。
具体实施方式
本发明发明人经过大量探索性实验,利用基因模块化方法开发出一种具备多功能(可注射性、自修复特性)的强力水下粘合蛋白材料,该新型多功能粘合材料在多个领域特别是生物医药领域将有广阔的应用前景,在此基础上完成了本发明。
本发明一方面提供一种融合蛋白,所述融合蛋白可以包括贻贝足丝蛋白片段和亮氨酸拉链片段。
本发明所提供的融合蛋白中,可以包括贻贝足丝蛋白片段,所述贻贝足丝蛋白(mussel foot proteins)可以选自包括但不限于紫贻贝足丝蛋白(mytilus edulis footprotein,mefp)、地中海贻贝足丝蛋白(Mytilus galloprovincialis foot protein)等,更具体可以选自包括但不限于mefp-3、mefp-5、Mgfp-3、Mgfp-5、mfp-3s、mfp3s-pep等或它们的变体,例如,所述贻贝足丝蛋白片段可以是:a)氨基酸序列如SEQ ID No.4-9其中之一所示的多肽片段,也可以是b)氨基酸序列与SEQ ID No.4-9其中之一具有80%以上、85%以上、90%以上、93%以上、95%以上、97%以上、或99%以上同源性、且具有a)限定的多肽片段的功能的多肽片段,所述b)中的氨基酸序列具体指:如SEQ ID No.4-9其中之一所示的氨基酸序列经过取代、缺失或者添加一个或多个(具体可以是1-50、1-30个、1-20个、1-10个、1-5个、或1-3个)氨基酸而得到的,或者在N-末端和/或C-末端添加一个或多个(具体可以是1-50个、1-30个、1-20个、1-10个、1-5个、或1-3个)氨基酸而得到的,且其编码的多肽片段具有如SEQ ID No.4-9其中之一所示的氨基酸序列所编码的多肽片段的功能的氨基酸序列。所述贻贝足丝蛋白通常来源于贻贝(拉丁名:Mytilidae),更具体可以来源于紫贻贝(mytilus eduli)、地中海贻贝(Mytilus galloprovinciali)等。
本发明所提供的融合蛋白中,可以包括卷曲螺旋结构(coiled-coiled)片段,所述卷曲螺旋结构片段通常能够使融合蛋白自组装呈现α-螺旋构象。所述卷曲螺旋结构片段可以是亮氨酸拉链片段、A1片段(例如,可以是SGDLXNXVAQLXRX(SEQ ID No.10)、VRSLXDXAAELXQX(SEQ ID No.11)、VSRLXNXIEDLXAXI(SEQ ID No.12)所示的氨基酸序列所编码的多肽片段,其中,X为E或K)、ZE/ZR片段、GCN4片段、γ52-88KI片段(例如,可以是SEQ IDNo.13所示的氨基酸序列所编码的多肽片段)、PCxP片段、NLP(Nucleoporin-likepolypeptide)片段等。在本发明一具体实施方式中,所述卷曲螺旋结构片段可以是亮氨酸拉链片段,所述亮氨酸拉链片段通常包括A片段、S片段和P片段。所述A片段可以是:c)氨基酸序列如SEQ ID No.1所示的多肽片段,也可以是d)氨基酸序列与SEQ ID No.1具有80%以上、85%以上、90%以上、93%以上、95%以上、97%以上、或99%以上同源性、且具有c)限定的多肽片段的功能的多肽片段。所述S片段可以是:e)氨基酸序列如SEQ ID No.2所示的多肽片段,也可以是f)氨基酸序列与SEQ ID No.2具有80%以上、85%以上、90%以上、93%以上、95%以上、97%以上、或99%以上同源性、且具有e)限定的多肽片段的功能的多肽片段。所述P片段可以是:g)氨基酸序列如SEQ ID No.3所示的多肽片段,也可以是h)氨基酸序列与SEQ ID No.3具有80%以上、85%以上、90%以上、93%以上、95%以上、97%以上、或99%以上同源性、且具有g)限定的多肽片段的功能的多肽片段。所述d)、f)、h)中的氨基酸序列具体指:分别如SEQ ID No.1、SEQ ID No.2、SEQ ID No.3所示的氨基酸序列经过取代、缺失或者添加一个或多个(具体可以是1-50、1-30个、1-20个、1-10个、1-5个、或1-3个)氨基酸而得到的,或者在N-末端和/或C-末端添加一个或多个(具体可以是1-50个、1-30个、1-20个、1-10个、1-5个、或1-3个)氨基酸而得到的,且其编码的多肽片段分别具有如SEQ ID No.1、SEQ ID No.2、SEQ ID No.3所示的氨基酸序列所编码的多肽片段的功能的氨基酸序列。在本发明一具体实施方式中,所述亮氨酸拉链片段自N端至C端依次包括A片段、S片段和P片段。
本发明所提供的融合蛋白中,贻贝足丝蛋白片段可以被插入亮氨酸拉链片段中。在本发明一具体实施方式中,所述融合蛋白自N端至C端依次包括A片段、S片段、贻贝足丝蛋白片段和P片段。
本发明所提供的融合蛋白中,所述融合蛋白可以是经酪氨酸酶(tyrosinase)修饰。所述经酪氨酸酶修饰通常指融合蛋白中至少部分的酪氨酸基团被转变为多巴基团(DOPA基团,3,4-二羟基苯丙氨酸),通常来说,酪氨酸基团的转化率可以为30%~80%、50%~65%、30%~35%、35%~40%、40%~45%、45%~50%、50%~55%、55%~60%、60%~65%、65%~70%、70%~75%或75%~80%。在本发明一具体实施方式中,对融合蛋白进行酪氨酸酶修饰时所使用的酪氨酸酶为SIGMA-ALDRICH试剂。
本发明所提供的融合蛋白中,还可以包括功能性蛋白片段,所述功能性蛋白片段可以是粘性蛋白片段、细胞粘附多肽片段、酶蛋白片段、生长因子片段、抗菌活性多肽片段、凝血因子片段、生物相容性多肽片段、抗生肽片段、监测多肽片段等中的一种或多种的组合,所述粘性蛋白片段可以是例如Mfp3s片段、MFPfast片段等,所述细胞粘附多肽片段可以是例如RGD片段(参见2.Heilshorn,S.C.;Liu,J.C.;Tirrell,D.A.,Cell-binding domaincontext affects cell behavior on engineered proteins.Biomacromolecules 2005,6(1),318-323.)、REDV片段(参见Girotti,A.;Reguera,J.;Rodríguezcabello,J.C.;Arias,F.J.;Alonso,M.;Matestera,A.,Design and bioproduction of a recombinant multi(bio)functional elastin-like protein polymer containing cell adhesionsequences for tissue engineering purposes.Journal of Materials ScienceMaterials in Medicine 2004,15(4),479-484.)等,所述酶蛋白片段(例如,有毒物质降解酶)可以是例如AdhD片段(参见Wheeldon,I.R.;Campbell,E.;Banta,S.,A ChimericFusion Protein Engineered with Disparate Functionalities—Enzymatic Activityand Self–assembly.Journal of Molecular Biology 2009,392(1),129-142.)、OPH片段(参见Girotti,A.;Reguera,J.;Rodríguezcabello,J.C.;Arias,F.J.;Alonso,M.;Matestera,A.,Design and bioproduction of a recombinant multi(bio)functionalelastin-like protein polymer containing cell adhesion sequences for tissueengineering purposes.Journal of Materials Science Materials in Medicine 2004,15(4),479-484.)等,所述生长因子片段可以是例如VEGF片段(参见Zisch,A.H.;Schenk,U.;Schense,J.C.;Sakiyama-Elbert,S.E.;Hubbell,J.A.,Covalently conjugated VEGF–fibrin matrices for endothelialization.Journal of Controlled Release 2001,72(1),101-113.)、KGF片段(参见Koria,P.;Yagi,H.;Kitagawa,Y.;Megeed,Z.;Nahmias,Y.;Sheridan,R.;Yarmush,M.L.,Self-assembling elastin-like peptides growth factorchimeric nanoparticles for the treatment of chronic wounds.Proceedings of theNational Academy of Sciences of the United States of America 2011,108(3),1034-1039.)等,所述抗菌活性多肽片段可以是例如HNP-2,4片段(参见Gomes,S.C.;Leonor,I.B.;Mano,J.F.;Rui,L.R.;Kaplan,D.L.,Antimicrobial functionalizedgenetically engineered spider silk.Biomaterials 2011,32(18),4255-4266.)、Hepcidin片段(参见Gomes,S.C.;Leonor,I.B.;Mano,J.F.;Rui,L.R.;Kaplan,D.L.,Antimicrobial functionalized genetically engineered spider silk.Biomaterials2011,32(18),4255-4266.)、Indolicidin片段(参见Tsai,C.W.;Hsu,N.Y.;Wang,C.H.;Lu,C.Y.;Chang,Y.;Tsai,H.H.;Ruaan,R.C.,Coupling molecular dynamics simulationswith experiments for the rational design of indolicidin-analogousantimicrobial peptides.Journal of Molecular Biology 2009,392(3),837-854.)、Plectasin片段(参见Zakeri,B.;Lu,T.K.,Synthetic Biology of AntimicrobialDiscovery.Acs Synthetic Biology 2013,2(7),358-372.)、β-defensin-1片段(参见Zakeri,B.;Lu,T.K.,Synthetic Biology of Antimicrobial Discovery.Acs SyntheticBiology 2013,2(7),358-372.)等,所述凝血因子片段可以是例如凝血III因子片段(参见Edgington,T.S.;Mackman,N.;Brand,K.;Ruf,W.,The structural biology ofexpression and function of tissue factor.Thrombosis&Haemostasis 1991,66(1),67.)、凝血V因子片段(参见Dahlback,B.;Hildebrand,B.,Inherited Resistance toActivated Protein C is Corrected by Anticoagulant Cofactor Activity Found tobe a Property of Factor V.Proceedings of the National Academy of Sciences ofthe United States of America 1994,91(4),1396-1400.)、凝血VII因子片段(参见O'Hara,P.J.;Grant,F.J.;Haldeman,B.A.;Gray,C.L.;Insley,M.Y.;Hagen,F.S.;Murray,M.J.,Nucleotide sequence of the gene coding for human factor VII,a vitamin K-dependent protein participating in blood coagulation.Proceedings of theNational Academy of Sciences of the United States of America 1987,84(15),5158-5162.)等,所述生物相容性多肽片段通常指具有提升融合蛋白生物相容性的多肽片段,所述抗生肽(antibacteialpeptides)片段通常指具有抑制微生物生长和/或杀死微生物功能的多肽片段,所述监测多肽片段可以是例如spytag、snooptag等。在本发明一具体实施方式中,所述功能性蛋白片段为spytag和/或snooptag,所述功能性蛋白片段的氨基酸序列如SEQ ID No.14和SEQ ID No.15所示,从而可以和融合有Spycatcher-(氨基酸序列如SEQ ID No.16所示)和Snoopcatcher(氨基酸序列如SEQ ID No.17所示)的融合蛋白反应。在本发明另一具体实施方式中,所述融合蛋白自N端至C端依次包括A片段、S片段、功能性蛋白片段、贻贝足丝蛋白片段和P片段。
本发明另一方面提供一种分离的多核苷酸,所述多核苷酸编码如上所述的融合蛋白。
本发明另一方面提供一种构建体,所述构建体含有如上所述的分离的多核苷酸。通过如上所述的分离的多核苷酸,构建所述构建体的方法对于本领域技术人员来说应该是已知的,例如,所述构建体可以由所述分离的多核苷酸插入到表达载体的多克隆位点构建而成,本领域技术人员可选择合适的表达载体用于构建所述构建体,例如,所述表达载体可以是细菌质粒等载体,再例如,所述表达载体可以是大肠杆菌质粒等,在本发明一具体实施方式中,所述表达载体可以是pHis-FUS质粒等。
本发明另一方面提供一种表达系统,所述表达系统含有如上所述构建体或基因组中整合有外源的如上所述的多核苷酸。所述表达系统通常可以是宿主细胞,任何适用于所述构建体进行表达的细胞都可以作为宿主细胞,例如,所述宿主细胞可以是原核细胞等,再例如,所述宿主细胞可以是细菌细胞等,在本发明一具体实施方式中,所述宿主细胞可以是例如大肠杆菌细胞等。
本发明另一方面提供所述融合蛋白的制备方法,包括:在适合表达所述融合蛋白的条件下,培养如上所述的表达系统。本领域技术人员可以根据所使用的宿主细胞的种类等,选择合适的培养条件。例如,培养中所用的培养基可以为各种常规培养基,本领域技术人员可根据经验选择适用的培养基,在适于宿主细胞生长的条件下进行培养。再例如,当宿主细胞生长到适当的细胞密度后,用合适的方法(如温度转换或化学诱导)诱导选择的启动子,将细胞再培养一段时间。通常来说,如上所述的融合蛋白可以在细胞内、或在细胞膜上表达、或分泌到细胞外,如果需要还可利用其物理的、化学的和其它特性通过各种分离方法分离和纯化重组的蛋白,这些方法对于本领域技术人员来说都应该是已知的,例如,可以采用复性处理、用蛋白沉淀剂处理(盐析方法)、离心、渗透破菌、超处理、超离心、分子筛层析(凝胶过滤)、吸附层析、离子交换层析、高效液相层析(HPLC)和其它各种液相层析技术等方法。
本发明所提供的融合蛋白的制备方法中,还可以包括:将表达所得的融合蛋白进行酪氨酸酶修饰。对化合物(例如,如上所述的融合蛋白)进行酪氨酸修饰的方法对本领域技术人员来说应该是已知的,例如,可以使所述融合蛋白处于酪氨酸酶存在的条件下。
本发明另一方面提供一种自组装胶体溶液,所述溶液包括如上所述的融合蛋白。所述自组装胶体溶液通常可以为溶液形式(例如,水溶液形式),可以在合适的条件下形成胶体。本领域技术人员可以选择合适种类和用量的溶剂与所述融合蛋白混合以形成所述自组装胶体的溶液,例如,可以是PBS溶液(例如,Shanghai ABCONE Biotechnology Co所提供的PBS缓冲液)等,再例如,本领域技术人员可以根据所制备的胶体中融合蛋白的含量适当调整溶液中融合蛋白的浓度。
本发明另一方面提供一种胶体,所述胶体包括如上所述的融合蛋白。所述胶体通常为水凝胶,即以水为分散介质的凝胶。所述胶体可以由如上所述的自组装胶体溶液制备获得,其主要是利用其中融合蛋白的自组装,从而获得了胶体。制备获得的胶体中,融合蛋白的含量通常可以为70mg/mL~250mg/mL。
本发明所提供的自组装胶体溶液和/或胶体中,可以是单组分的,即,仅含有一种如上所述的融合蛋白,也可以是多组分的,即含有两种以上如上所述的融合蛋白。在本发明一具体实施方式中,所述自组装胶体溶液和/或胶体中,其所包含的融合蛋白中,其贻贝足丝蛋白可以是mefp-3和mefp-5的组合,例如,可以包括含有mefp-3肽段的融合蛋白以及含有mefp-5肽段的融合蛋白,两者的比例可以是1:0.01-100、1:0.1-10、1:0.1-0.3、1:0.3-0.5、1:0.5-1、1:1-2、1:2-3、1:3-5或1:5-10。
本发明另一方面提供如上所述的融合蛋白、自组装胶体溶液、胶体在水下粘合剂制备领域的用途,更具体可以是一种仿生水下粘合剂材料。
本发明提供包含贻贝类足丝蛋白与卷曲螺旋结构蛋白或蛋白结构域的融合蛋白,运用分子生物学技术成功构建重组质粒,将其转入大肠杆菌后表达重组蛋白,并通过聚丙烯酰胺凝胶电泳及免疫印迹实验鉴定蛋白的表达,通过圆二色谱实验分析融合蛋白对重组蛋白二级结构的影响。为了增强蛋白的界面粘性,通过酪氨酸酶对融合蛋白进行修饰,并用NBT染色实验验证。通过原子力显微镜对混合蛋白分别在酪氨酸酶修饰和未修饰条件下的形貌进行表征,以比较各种蛋白对于云母片的吸附量以及自身团聚能力。此外,本发明还进一步通过融合蛋白制备水凝胶,通过使用万能材料试验机搭接剪切测试水凝胶的粘合力,发现蛋白水凝胶在空气湿度为30%条件下中测试的最大粘合力强度可以达到286kPa,在水环境条件下可以达到近100kPa,分别为对照蛋白的8.8倍和16倍。此外,还通过搭接剪切测试、扫描电子显微镜及流变学测试实验验证了蛋白水凝胶粘合力显著提升的原因,材料整体粘合力的增强来源于其界面粘性和内在粘性的同时提升。进一步的,本发明发明人还发现,经酪氨酸酶修饰后A-S-Spytag-Mefp3-P与A-S-Snooptag-Mefp5-P靠近后的界面在荧光显微镜下展示的变化过程,从而发现A35 MO内A-S-Mefp3-P和A-S-Mefp5-P存在着相互作用。可见,通过对水凝胶材料性质的自修复实验、3D打印实验及对骨骼、玻璃片的水下粘合效果测试实验,证明了其具备自修复性及可注射性,以及水下粘合的具体可实施性,体现出其良好的应用前景。
综上所述,本发明将贻贝足丝蛋白与卷曲螺旋结构进行基因重组,从而提供了一种新的融合蛋白,所述融合蛋白可以在溶液中自组装形成凝胶材料,兼具贻贝足丝蛋白的界面粘性以及卷曲螺旋结构的内在粘性,从而实现水下超粘合强度。与现有技术相比,本发明同时增强材料的界面粘性与内在粘性从而提升整体粘合力,得到一种具有自修复性和可注射性的多功能水下粘合材料,探索出构建水下粘合材料的新策略,并通过实例表明该材料潜在的应用性。
以下通过特定的具体实例说明本发明的实施方式,本领域技术人员可由本说明书所揭露的内容轻易地了解本发明的其他优点与功效。本发明还可以通过另外不同的具体实施方式加以实施或应用,本说明书中的各项细节也可以基于不同观点与应用,在没有背离本发明的精神下进行各种修饰或改变。
在进一步描述本发明具体实施方式之前,应理解,本发明的保护范围不局限于下述特定的具体实施方案;还应当理解,本发明实施例中使用的术语是为了描述特定的具体实施方案,而不是为了限制本发明的保护范围;在本发明说明书和权利要求书中,除非文中另外明确指出,单数形式“一个”、“一”和“这个”包括复数形式。
当实施例给出数值范围时,应理解,除非本发明另有说明,每个数值范围的两个端点以及两个端点之间任何一个数值均可选用。除非另外定义,本发明中使用的所有技术和科学术语与本技术领域技术人员通常理解的意义相同。除实施例中使用的具体方法、设备、材料外,根据本技术领域的技术人员对现有技术的掌握及本发明的记载,还可以使用与本发明实施例中所述的方法、设备、材料相似或等同的现有技术的任何方法、设备和材料来实现本发明。
除非另外说明,本发明中所公开的实验方法、检测方法、制备方法均采用本技术领域常规的分子生物学、生物化学、染色质结构和分析、分析化学、细胞培养、重组DNA技术及相关领域的常规技术。这些技术在现有文献中已有完善说明,具体可参见Sambrook等MOLECULAR CLONING:A LABORATORY MANUAL,Second edition,Cold Spring HarborLaboratory Press,1989 and Third edition,2001;Ausubel等,CURRENT PROTOCOLS INMOLECULAR BIOLOGY,John Wiley&Sons,New York,1987 and periodic updates;theseries METHODS IN ENZYMOLOGY,Academic Press,San Diego;Wolffe,CHROMATINSTRUCTURE AND FUNCTION,Third edition,Academic Press,San Diego,1998;METHODS INENZYMOLOGY,Vol.304,Chromatin(P.M.Wassarman and A.P.Wolffe,eds.),AcademicPress,San Diego,1999;和METHODS IN MOLECULAR BIOLOGY,Vol.119,ChromatinProtocols(P.B.Becker,ed.)Humana Press,Totowa,1999等。
本发明实施例中,如无特殊说明,酶连反应体系(10μL)如表1所示:
表1
本发明实施例中,如无特殊说明,PCR扩增及PCR鉴定采用的反应体系(50μL)包括以下成份:
PCR反应条件为:98℃预变性20s,98℃10s,58℃30s,72℃45s,35个循环,最后72℃延伸5min,以去离子水为阴性对照。
本发明实施例中,如无特殊说明,使用的LB培养基购自(台湾生工,L001-1kg),抗生素溶液的配制方法为:羧苄青霉素(macklin,C805408)配制成100mg/ml的水溶液于-20℃保存,使用时抗生素终浓度为50ng/ml。
本发明实施例中,如无特殊说明,使用的裂解液,清洗液,洗脱液,透析液配方如下:
(1)裂解液:8M盐酸胍水溶液
(2)清洗液:20mM咪唑水溶液
(3)洗脱液:1000mM咪唑水溶液
(4)透析液:dd H2O
本发明实施例中所使用的试剂及耗材如表2所示:
表2
本发明实施例中所用到的大肠杆菌BL21(DE3),可从TransGene Biotech(Beijing)购买,目录号为CD601。
本发明实施例中所用到的pHis-FUS(LC)质粒参照(Cell-free Formation of RNAGranules:Low Complexity Sequence Domains Form Dynamic Fibers withinHydrogels.2012,Cell,Volume 149,Issue 4,11May 2012,Pages 753–767.Masato Kato,Tina W.Han,Steven L.McKnight),美国西南医学院的McKnight教授课题组已将上述质粒馈赠给本申请人。
实施例1
功能性A-S-P融合蛋白的构建、表达及提纯:
构建A-S-P重组质粒,以实验室现有的pHis-FUS(LC)质粒为基础,插入目的基因片段。A的氨基酸序列如SEQ ID NO:1所示,S的氨基酸序列如SEQ ID NO:2所示,P的氨基酸序列如SEQ ID NO:3所示,A-S-P的氨基酸序列如SEQ ID NO:20所示。构建上述所需的表达质粒,其涉及的主要步骤包括:
(1)将S片段插入载体pHis-FUS(LC):其中S蛋白基因片段由苏州金唯智公司合成,将其插入至pHis-FUS(LC)质粒,得到载体pHis-S-FUS(LC),S前后两端的酶切位点为NcoI与BamHI。
(2)将A片段插入载体pHis-S-FUS(LC):利用引物扩增得到基因A片段(无需模板)。正向引物A-F的DNA序列见SEQ ID NO:21,反向引物A-R的DNA序列见SEQ ID NO:22。载体pHis-S-FUS(LC)与PCR获得的基因片段A分别用限制性内切酶NdeI与NcoI进行酶切实验,温度为37℃,时间为1h。酶切完毕后,利用北京全式金生物技术有限公司生产的试剂盒EasyPure Quick Gel Extraction Kit进行酶切产物的胶回收。将回收产物用T4连接酶连接,温度为25℃,时间为1h,得到载体A-S-FUS(LC)。
(3)将P片段插入载体A-S-FUS(LC):利用引物扩增得到基因P片段(无需模板),正向引物P-F和反向引物P-R的DNA序列详见附录。将载体A-S-FUS(LC)与PCR获得的基因片段P分别用限制性内切酶BamHI与XhoI酶切,所得的回收产物利用T4连接酶连接,得到目标载体A-S-P,质粒图谱见图3。
实施例2
功能性A-S-M-P蛋白的构建、表达与纯化:
构建A-S-M-P重组质粒,其中M为不同功能化基团的肽段(M分别为Mefp3、Mefp5、Spytag-Mefp3、Snooptag-Mefp5)。在BL21(DE3)大肠杆菌内转入A-S-M-P表达质粒,使其表达A-S-M-P融合蛋白。其中Mefp3的氨基酸序列见SEQ ID NO:4,Mefp5的氨基酸序列见SEQID NO:5,Spytag的氨基酸序列见SEQ ID NO:14,Snooptag的氨基酸序列见SEQ ID NO:15,A-S-Mefp3-P的DNA序列见SEQ ID NO:23,A-S-Mefp5-P的DNA序列见SEQ ID NO:26,A-S-Spytag-Mefp3-P的DNA序列见SEQ ID NO:29,A-S-Snooptag-Mefp5-P的DNA序列见SEQ IDNO:32。
以构建成功的A-S-P质粒为基础,插入目的基因片段。构建上述所需的表达质粒,其涉及的主要步骤包括:
(1)对于表达质粒A-S-Mefp3-P,A-S-Mefp5-P,A-S-Spytag-Mefp3-P以及A-S-Snooptag-Mefp5-P的构建,利用限制性内切酶HindIII与BamHI按照常规方法切割表达质粒A-S-P使质粒线性化,根据Mefp3,Mefp5,Spytag-Mefp3,Snooptag-Mefp5与酶切后的表达载体的基因序列设计适用于酶切连接的上下游特异引物。
(2)分别通过PCR方法扩增构建各质粒所用的Mefp3片段、Mefp5片段、Spytag-Mefp3片段和Snooptag-Mefp5片段。
对于构建质粒A-S-Mefp3-P:Mefp3片段的正向引物Mefp3-F的DNA序列见SEQ IDNO:24,反向引物Mefp3-R的DNA序列见SEQ ID NO:25。Mefp3的模板质粒来源于pET11d-CsgA-Mefp3,参考文献Strong underwater adhesives made by self-assembling multi-protein nanofibres.Nature Nanotechnology 2014,9(10),858-866.Zhong,C.;Gurry,T.;Cheng,A.A.;Downey,J.;Deng,Z.T.;Stultz,C.M.;Lu,T.K.,所构建的质粒图谱A-S-Mefp3-P见图4。
对于构建质粒A-S-Mefp5-P:Mefp5片段的正向引物Mefp5-F的DNA序列见SEQ IDNO:27,反向引物Mefp5-R的DNA序列见SEQ ID NO:28。Mefp5的模板质粒来源于pET11d-Mefp5-CsgA,参考文献Strong underwater adhesives made by self-assembling multi-protein nanofibres.Nature Nanotechnology 2014,9(10),858-866.Zhong,C.;Gurry,T.;Cheng,A.A.;Downey,J.;Deng,Z.T.;Stultz,C.M.;Lu,T.K.,所构建的质粒图谱A-S-Mefp5-P见图5。
对于构建质粒A-S-Spytag-Mefp3-P:Spytag-Mefp3片段的正向引物Spytag-Mefp3-F的DNA序列见SEQ ID NO:30,反向引物Spytag-Mefp3-R的DNA序列见SEQ ID NO:31。所构建的质粒图谱A-S-Spytag-Mefp3-P见图6。
对于构建质粒A-S-Snooptag-Mefp5-P:Snooptag-Mefp5片段的正向引物Snooptag-Mefp5-F的DNA序列见SEQ ID NO:33,反向引物Snooptag-Mefp5-R的DNA序列见SEQ ID NO:34。所构建的质粒图谱A-S-Snooptag-Mefp5-P见图7。
将PCR扩增所得的目标片段(Mefp3片段、Mefp5片段、Spytag-Mefp3片段和Snooptag-Mefp5片段)分别用限制性内切酶HindIII与BamHI按照常规酶切方法切割,并分别与上述的经限制性内切酶HindIII与BamHI酶切并线性化后的A-S-P载体,用T4酶连方法得到目标表达载体,根据各片段的长度和浓度,按所需比例分别加至EP管中。反应体系在16℃中恒温孵育一夜。
重组质粒的转化步骤如下:
(1)将A-S-P以及A-S-Mefp3-P、A-S-Mefp5-P、A-S-Spytag-Mefp3-P、A-S-Snooptag-Mefp5-P的10μL连接产物分别转化于大肠杆菌克隆菌株DH5α(康为世纪CW08085)具体步骤如下:
a)取含100μL DH5α感受态细胞离心管,静置于冰盒降温融化,待其解冻时,加入10μL连接产物,轻弹混匀,置于冰盒30min。
b)42℃热激45s,迅速将离心管转移到冰盒中,静置2min。
c)向离心管内加入500μL无菌LB培养基,混匀后在37℃条件下用摇床振荡培养30min。
d)将步骤c中已经孵育30min的菌液在1000×g条件下离心2min,除去上清约400μL,将剩余100μL左右液体吹吸均匀,滴入含50μg/mL羧苄青霉素的LB固体培养基上,用涂布棒将细菌均匀涂开,至菌液被平板完全吸收,用恒温培养箱在37℃条件下倒置培养12h左右。
(2)挑选单克隆,并接种于含50μg/mL带羧苄青霉素抗性的LB液体培养基中,利用菌落PCR鉴定是否含有目的基因片段,如扩出条带则证明质粒转化成功。采用提质粒试剂盒(天根DP103)按操作说明提取质粒,由苏州金唯智公司测序后,进一步鉴定重组质粒。若测序结果正确,则将重组质粒转入大肠杆菌感受态BL21(DE3)(TransGene Biotech CD601)中,详细步骤如下:
a)取含100μL大肠杆菌感受态细胞BL21(DE3)离心管,静置于冰盒。
b)待BL21感受态细胞解冻,加入约1μL所提取质粒,用移液枪轻吹混匀,置于冰盒30min。
c)42℃热激45s,立刻将离心管转移到冰盒中,静置于冰盒2min。
d)向离心管内加入500μL无菌LB培养基(无抗生素添加),混匀后将离心管置于50mL离心管,在37℃条件下用摇床振荡培养30min。
e)将步骤d中已经孵育30min的菌液在1000×g条件下离心2min,除去上清约400μL,将剩余100μL左右液体吹吸均匀,滴入含50μg/mL羧苄青霉素的LB固体培养基上,用涂布棒将细菌均匀涂开,至菌液被平板完全吸收,在37℃恒温培养箱倒置培养12h左右。
重组A-S-M-P蛋白的诱导表达及提纯步骤如下:
(1)挑取含有目标质粒(A-S-P,A-S-Mefp3-P,A-S-Mefp5-P,A-S-Spytag-Mefp3-P以及A-S-Snooptag-Mefp5-P)的BL21(DE3)大肠杆菌接种到100mL含有50μg/mL羧苄青霉素抗性的LB液体培养基中,于37℃摇床振荡培养过夜,(约12h),摇床转速为220rpm。
(2)取10mL过夜培养的菌液按1:100比例接种到1L含50μg/mL羧苄青霉素的LB液体培养基中37℃摇荡培养至OD600在0.8-1.0之间。
(3)加入1mL 0.5 M IPTG到(2)所述菌液中,于37℃摇床摇荡诱导3h。
(4)诱导后的菌液离心收集菌体,称取菌体质量置于-80℃冷藏。
(5)每克菌体加入10mL 8M盐酸胍溶液,搅拌12h。
(6)21000×g离心1h去除菌体沉淀保留上清,在上清中加入Ni-NTA柱子(GeneralElectric Company,货号:17057501)并置于旋转摇床结合30min。
(7)利用重力层析柱(BBI,镍柱)提取蛋白,将上述蛋白溶液加入到重力层析柱,并用200-300mL清洗液(20mM咪唑清洗液)清洗重力层析柱去除杂蛋白。
(8)用20-30mL洗脱液(1000mM咪唑清洗液)在重力层析柱中洗脱重力层析柱上结合的目标蛋白(A-S-P,A-S-Mefp3-P,A-S-Mefp5-P,A-S-Spytag-Mefp3-P以及A-S-Snooptag-Mefp5-P),并收集流下的目标蛋白溶液,利用Nanodrop 2000测定蛋白浓度。
实施例3
融合蛋白A-S-M-P的表达验证:
通过考马斯亮蓝染色和蛋白免疫印迹实验比对蛋白标记物进行验证,检验融合蛋白是否能够成功表达。
蛋白聚丙烯酰胺凝胶电泳主要步骤如下:
(1)取90μL洗脱的目标蛋白与30μL 4×上样缓冲液均匀混合;
(2)加20μL混合液滴加至蛋白胶泳道中,并加入标准蛋白标记物作为对照;
(3)将蛋白胶置于电泳缓冲液中,在165V电压下采用电泳方式45min进行分离;
(4)将蛋白胶取出后用考马斯亮蓝溶液染色1h后进行脱色(脱色液:50%ddH2O,40%甲醇,10%乙酸);
(5)脱色进行1h后,用蛋白胶扫描仪进行拍照。
蛋白免疫印迹鉴定实验主要步骤如下:
(1)同蛋白聚丙烯酰胺凝胶电泳步骤(1)~(3);
(2)使用蛋白胶电转仪将蛋白胶转印到硝酸纤维素薄膜上;
(3)将膜浸至含20%的脱脂奶粉溶液中,振荡孵育1h,之后使用TBST溶液中荡洗3次,每次进行10min;
(4)将膜浸至20mL包含经稀释5000倍一抗原液的TBST溶液中,振荡孵育1h,之后使用TBST溶液荡洗荡洗3次,每次进行10min;
(5)将膜浸至20mL包含经稀释2000倍的二抗原液的TBST溶液中,振荡孵育1h,之后使用TBST溶液荡洗荡洗3次,每次进行10min;
(6)对膜进行显色操作,之后使用蛋白胶扫描仪进行拍照。
所得结果如图8所示,将图8b所示标准蛋白标记物作为参照,可知图8a和8c中显示A-S-P,A-S-Mefp3-P,A-S-Mefp5-P蛋白分别在30~40kD、30~40kD和20~30kD出现了相对应的目标条带。(图中AS3P、AS5P、ASP分别代表A-S-Mefp3-P、A-S-Mefp5-P、A-S-P蛋白样品)对应的目标条带比理论分子量略微偏大,这可能是因为A-S-P系列蛋白有较多的疏水基团,造成了蛋白在电泳中移动的速率比一般的蛋白移动稍慢,因而出现在理论位置的上方,符合经验值;同时,所有表达的蛋白C端都带有6个组氨酸标签,根据抗原与抗体相互作用的原理,通过蛋白免疫印迹鉴定试验结果所示,在相应位置出现了目标条带。综合上述说明,纯化的蛋白即是目标蛋白,从而验证了蛋白的成功表达与纯化。
实施例4
A-S-P、A-S-M-P二级结构的表征:
利用圆二色谱对A-S-P、A-S-Mefp3-P、A-S-Mefp5-P二级结构进行表征,检验融合蛋白的成分Mefp-3、Mefp-5是否会对分子的构象带来极大改变。主要步骤如下:
(1)将新制的蛋白溶液利用去离子水进行多次透析。
(2)取100μL的目标蛋白溶液,利用圆二色谱仪进行圆二色谱分析,测量波长为190-250nm。实验使用Chirascan spectrometer(Applied Photophysics)仪器进行测试。
根据圆二色谱谱图9所示,三种蛋白样品均在208nm附近有一个负峰,在223nm附近有一个负峰,两者都属于α-helix的特征峰,证明A-S-P系列蛋白均含α-helix的二级结构3;同时根据图9b各二级结构所占的百分比结果显示,A-S-P系列蛋白中含有比例相近的二级结构,这说明本实验所融合的Mefp-3和Mefp-5蛋白对A-S-P的二级结构没有显著的影响。以上表明,同A-S-P蛋白相似,A-S-Mefp3-P和A-S-Mefp5-P中也存在α-helix的二级结构,由于Mefp-3和Mefp-5是无规结构,则α-helix主要来源于A和P片段,可推测它们的成胶方式与A-S-P类似,都源自于其自组装能力。
实施例5
重组蛋白A-S-M-P酪氨酸酶的修饰及验证:
通过对融合蛋白A-S-Mefp3-P,A-S-Mefp5-P,A-S-Spytag-Mefp3-P以及A-S-Snooptag-Mefp5-P进行酪氨酸酶修饰,使其中的酪氨酸转变为多巴成分,以期水凝胶的界面粘性由于多巴的生成得以增强。
酪氨酸酶修饰的主要步骤如下:
(1)蛋白提取过程如实施例2中“重组A-S-M-P蛋白的诱导表达及提纯步骤”所述,待目标蛋白与镍柱结合后,沉积在重力层析柱上,利用100-200mL 20mM咪唑缓冲液清洗杂蛋白。
(2)用20mL PBS缓冲液清洗已经去除杂蛋白后的层析柱;
(3)将4mL 0.5mg/mL酪氨酸酶溶液(配方:20mM硼酸钠,100mM PBS缓冲液,抗坏血酸100mM,pH=7.0)加入层析柱中,置于旋转摇床在室温下反应2h;
(4)加入20mL PBS缓冲液清洗酪氨酸酶;
(5)加入20-30mL洗脱液(400mM咪唑缓冲液)在重力层析柱中洗脱重力层析柱上结合的目标蛋白(经酪氨酸酶修饰后的A-S-Mefp3-P,A-S-Mefp5-P,A-S-Spytag-Mefp3-P以及A-S-Snooptag-Mefp5-P),并收集流下的目标蛋白溶液,利用Nanodrop 2000测定蛋白浓度。
为了验证A-S-Mefp3-P和A-S-Mefp5-P蛋白是否被酪氨酸酶成功修饰使其中的酪氨酸转变为二羟苯丙氨酸,对修饰后的蛋白样品进行NBT染色实验表征4。氯化硝基四氮唑蓝(NBT)是用于鉴定蛋白中的酪氨酸是否转变成了邻苯二酚或醌类结构的特征染料,在还原剂的作用下能生成蓝色化合物,通过该染色反应证明邻苯二酚结构,即多巴是否成功被催化引入。主要步骤如下:
(1)将经酪氨酸酶修饰和未经修饰的蛋白溶液,通过点转仪转印于硝酸纤维素薄膜;
(2)将膜浸至20mL新制0.6mg/mL的NBT溶液(2M磷酸钾和甘氨酸缓冲液,pH=10.0)中,在避光条件下静置孵育45min;
(3)用10mL 0.16M硼酸钠溶液清洗两次,浸至20mL的硼酸钠溶液中过夜脱色,之后进行拍照。
经酪氨酸酶修饰后的蛋白NBT染色实验结果如图10所示。经酪氨酸酶修饰后的蛋白在和氯化硝基四氮唑蓝作用之后变成了蓝色,这证明经酪氨酸酶修饰后的A-S-Mefp3-P和A-S-Mefp5-P蛋白中部分的酪氨酸变为多巴结构,A-S-Mefp3-P和A-S-Mefp5-P蛋白样品成功地被酪氨酸酶修饰;而未经修饰的A-S-Mefp3-P和A-S-Mefp5-P蛋白没有显示出蓝色,验证出它们不含多巴结构。
实施例6
原子力显微镜(AFM)形貌表征实验:
利用所得的目标蛋白溶液进行原子力显微镜(AFM)表面形貌表征,主要步骤如下:
(1)将新制的浓度为1μM左右的目标蛋白溶液(4mg/mL)滴加到干净平整的云母片表面,孵育12h后用去离子水冲洗云母片表面并用氮气吹干;
(2)用原子力显微镜在轻敲模式下进行形貌扫描。所用微型探针型号为AC 160(k=26.1N/m,ν~300kHz).实验使用Asylum MFP-3D AFM(Asylum Research)仪器进行测试。
得到的图像如图11显示。从整体上看,A-S-P系列蛋白均形成蛋白团聚块,未出现特殊的形貌。在等质量的所有种类蛋白溶液样品中,A-S-P蛋白团聚物的体积较小,且在云母表面吸附量较少。通过A-S-Mefp3-P、A-S-Mefp5-P和等质量A-S-Mefp5-P与A-S-Mefp3-P混合溶液以及它们分别经酪氨酸修饰后对应的溶液比照,可发现经过酪氨酸酶修饰后的蛋白团聚更紧密,且在云母表面的吸附量均高于其未经酪氨酸酶修饰的蛋白样品。可以推测,在亮氨酸拉链结构域上增添粘性蛋白Mefp-3和Mefp-5,不会改变原亮氨酸拉链蛋白的形貌,同时能够增强对云母表面吸附的能力;而经酪氨酸酶修饰后的样品增强了蛋白团聚的能力,此外进一步增强对云母表面的吸附能力,这可能是多巴的粘性特征带来的影响。同时能够发现其中团聚能力最明显加强的是经酪氨酸酶修饰后等质量A-S-Mefp5-P与A-S-Mefp3-P,这可能是由于经修饰后的Mefp-5和Mefp-3之间的相互作用所带来的蛋白内在粘性提升所致。
实施例7
A-S-P系列蛋白成胶实验:
通过使用冻干后滴加PBS溶液方法,无需添加化学交联剂,A-S-P系列蛋白即可自组装成水凝胶胶,主要步骤如下:
(1)将提纯后的蛋白溶液样品置于透析袋内,并将其置于去离子水中透析3次,每次透析30min;
(2)将透析后的样品装入50mL离心管内,并置于-80℃冰箱内冷冻;
(3)取出完全冷冻后的样品,打开离心管盖子,用封口膜封口,并用针头在膜上开口,保证气体的流通;
(4)将离心管置于冷冻真空干燥机24h;
(5)将样品取出,称重,计算指定浓度下样品的体积,滴加100mM PBS溶液至所需体积;
(6)将样品静置3h可形成特定浓度的自组装水凝胶。
如图12所示为A-S-P系列蛋白成水凝胶状态后的图片。其中A-S-P蛋白所成水凝胶(简称ASP)无色透明,未经修饰的A-S-Mefp3-P、A-S-Mefp5-P、等质量A-S-Mefp3-P与A-S-Mefp5-P混合蛋白(分别简称为A3、A5、A35)略带淡黄色;而经过酪氨酸酶修饰后的A-S-Mefp3-P、A-S-Mefp5-P、等质量A-S-Mefp3-P与A-S-Mefp5-P混合蛋白(分别简称为A3MO、A5MO、A35 MO)颜色略带褐色,这可能是由于其中部分多巴氧化成醌的结果所致。从成胶形态来看,ASP系列的状态大致相同。而其性能,如粘弹性及粘性强度,需要进一步通过对流变仪测试及万能材料试验机测试结果进行分析。
实施例8
A-S-P系列蛋白胶的粘合强度测试:
万能材料试验机测试粘性,主要步骤如下:
(1)称取3mg 250mg/mL水凝胶样品,将其涂抹于不锈钢片(尺寸为10mm×50mm×0.025mm,涂抹范围10mm×10mm),并用相同的不锈钢片粘合于涂抹部分。
(2)将样品置于37℃培养箱中孵育2h。
(3)取样品,使用万能材料试验机进行测试,室温条件,湿度为30%。设定为5mm/min,两夹具间距为60mm。
(4)对于在水中测试的样品,取样后室温条件下浸至水中5min,样品处于水环境下使用万能材料试验机进行测试。其他仪器设置条件与(3)相同。
如图13所示,通过基因模块化构建添加的粘性蛋白Mefp-3、Mefp-5后的A3和A5粘合强度比ASP明显提高,分别是ASP的2倍和3.1倍。而A3 MO和A5 MO粘合强度分别是相应未经修饰的水凝胶的2倍和1.7倍,分别是ASP的4.1倍和5.2倍。而A35 MO粘合强度高达286.14±24.37kPa,是相应未经修饰水凝胶的1.7倍,是ASP粘合强度的8.8倍。从以上实验数据进行分析可得出:
(1)经过增加粘性蛋白后的A3和A5相较ASP提高了粘合强度。这是由于粘性蛋白对界面粘合起到增强的作用。
(2)经过酪氨酸酶修饰后所成的A3 MO和A5 MO相比ASP以较大幅度增大了粘合强度。这是由于经过酪氨酸酶修饰后所形成的的多巴成分由于氢键、金属配位作用对界面粘合有了更进一步增强的效果。而经酪氨酸酶修饰后所成的A5 MO粘合强度高于A3 MO,可能的原因是,Mefp-5中含有更多的酪氨酸和更多的赖氨酸,呈现出更强的界面粘性。这些结果与先前研究结果所述Mfp-5比Mfp-3的粘合能力更强相一致。
(3)经酪氨酸酶修饰后所成的A35 MO粘合强度相较ASP有了最大的提高效果。从A3MO和A5 MO的粘合强度数据看来,A5 MO的粘合强度高于A3 MO,似乎将二者混合后制备的样品粘合强度应介于二者之间。然而实际得到的粘合强度却并非简单的二者叠加后的平均,而是超过了其中任意一种样品。因而可以推测,A35 MO粘合强度显著提升的原因可能来源于两方面:第一,经酪氨酸酶修饰后的A-S-Mefp3-P蛋白和A-S-Mefp5-P蛋白中的多巴成分促使其在界面粘性得到提升。不锈钢片成分中包含多种金属及其氧化物,其中金属能够与多巴形成配位键,金属氧化物中的氧能够与多巴生成氢键,从而增加界面粘性。第二,由于A-S-Mefp3-P和A-S-Mefp5-P中存在相互作用(来源于亮氨酸拉链的分子识别以及Mefp-3和Mefp-5之间的分子间氢键等作用)从而可能使其交联网络更为致密,引起内在粘性提升。由于A35 MO界面粘性与内在粘性相较ASP同时加强,这两方面的作用共同提升了水凝胶的粘合强度。
该实验成功地证明了,经酪氨酸酶修饰后的A-S-Mefp3-P和A-S-Mefp5-P蛋白等质量混合所成水凝胶具有最高的整体粘合强度,无论在未经酪氨酸酶修饰还是经修饰的条件下,均高于A-S-Mefp3-P、A-S-Mefp5-P在相同条件下的蛋白水凝胶。而在Lu课题组水下粘合材料的研究中,经过AFM力学测试的实验表明2,经融合表达大肠杆菌中淀粉样蛋白CsgA和贻贝足丝蛋白Mfp-3、Mfp-5而得的两种仍然具有自组装性质的粘性纳米纤维CsgA-Mfp3和Mfp5-CsgA被等摩尔比混合时,达到了最大的粘合效果,均高于CsgA-Mfp3和Mfp5-CsgA。可见,无论使用宏观层面(搭接剪切测试)还是微观层面(AFM)的实验方法,包含Mfp-3和Mfp-5的粘性融合蛋白混合后的粘性效果均显著增强,均强于其每种粘性融合蛋白单独存在时的效果。因此可以推测,混合后包含Mfp-3、Mfp-5的融合蛋白分子间发生了相互作用,加强了粘合特征,这一特性不仅在微观层面上体现,也在宏观层面上有所展示。
如图14所示,ASP和A35 MO水凝胶的粘合力分别比在同等孵育条件下空气中测得的粘合力降低约5.3倍和2.9倍,但A35 MO依然显示出较高的粘合力,达近100kPa,是ASP水下粘合力的16倍。同时可见,A35 MO在水下测得的粘合力比其在空气中测得的粘合力降低的幅度小于ASP,这说明A35 MO在水下粘合方面相较ASP有着更优越的表现。
为了验证A35 MO粘合力得到显著提升的原理,本发明对A35 MO静置十天后的水凝胶(以下简称为A35 MO10)进行搭接剪切测试,实验结果如图16所示。A35 MO10在室温21℃,湿度30%的空气中所测得的粘合力为80.94kPa,仅为A35 MO粘合力的29.16%。而在相应的水下测试中所得粘合力为23.64kPa,仅为A35 MO粘合力的24.22%。从以上数据可推测,经过酪氨酸酶修饰后的A35 MO中的多巴结构由于在空气中可被氧化为醌结构,大大降低了其界面粘性;从而使得A35 MO10与A35 MO相比,无论处于空气环境中还是水环境中测试,其粘合力都有明显的降低。但同时能够发现,A35 MO10的粘合力依然强于ASP水凝胶,这可能是由于A35 MO10中由于醌的生成所带来的内在粘性的提升。以上可以说明,A35 MO粘合力的显著增强一方面是来源于经过酪氨酸酶修饰后形成的多巴结构与界面的粘合作用。
流变仪测试流变性质,取500mL各凝胶样品,将其于流变仪cp25-2转子表面充分涂匀,并在测试之前使用刮勺除去周边多余部分。
(1)振幅扫描
设置条件:应变(形变)阶梯变化(CSD,对数坐标):应变幅度γ=0.01~100%,振荡角频率ω=10rad/s。
如图18所示,通过流变仪的振幅扫描模式测量ASP和A35 MO的线性粘弹区范围及角频率为10rad/s时的粘弹性。从应变幅度(Strain)来看,ASP线性粘弹区终点约为1%,而A35 MO则超过了10%。同时,可看出A35 MO的在线性粘弹区范围内储存模量(Storagemodulus)比ASP高出一个数量级。
由此可见,A35 MO相较ASP的韧性及弹性均提高。可以得出经酪氨酸酶修饰后的A-S-Mefp3-P和A-S-Mefp5-P混合水凝胶,相较ASP而言不仅发生了分子级别的改变,由于Mefp-3和Mefp-5的引入以及它们相互之间的作用,水凝胶整体更加富有韧性和弹性,这可能是由于A35 MO已经同原ASP的交联结构有明显变化,变得更为紧致,从而对于材料性能有了进一步地提高。此结论与图17所示的SEM实验所证明结论一致。
(2)频率扫描
设置条件:频率扫描(CSD,对数分布),振荡角频率ω=100~0.01rad/s,应变幅度γ=1%(需采用LVE范围内的应变,可从振幅扫描实验中获得)。
结果如图19所示。
为进一步研究A35 MO的结构性质,使用液氮冷冻水凝胶样品并置于冻干机静置的方法制备样品,并用SEM观察ASP和A35 MO样品的内部结构。主要步骤如下:
(1)将水凝胶样品使用液氮冷冻后,置于冻干机内48h。
(2)将冻干后的样品通过导电胶粘附到样品检测台,暴露出剖面,并镀金后进行成像表征,喷金仪型号为SBC-12。实验使用SUPRA 55SAPPHIRE扫描电子显微镜(Zeiss)进行测试,操作电压为2kV。
如图17所示,ASP经冷冻干燥后所得样品其内部孔径大小约为5.29μm,A35 MO内部孔径大小约为1.86μm。ASP样品的孔径大小约为A35 MO的3倍。由此可推测,A35 MO内蛋白分子所形成的交联程度大于ASP样品,形成的原因可能来自于经过酪氨酸酶修饰后的A-S-Mefp3-P和A-S-Mefp5-P蛋白中的多巴相互作用等其它作用,从而引起交联密度的增加。而交联密度的增加引起A35 MO的内在粘性的提升,从第二个层面解释了A35 MO粘合力加强的原因。
骨粘合实验,主要步骤如下:
(1)取5mg 250mg/mL水凝胶样品,将其涂于培养皿底部,涂胶面积为1cm×2cm,并在37℃培养箱中孵育15min。
(2)取10mg同种类水凝胶样品,将其均匀涂于牛骨底部表面,并在37℃培养箱中孵育1h。
(3)将20mL去离子水注入培养皿内。
(4)拎起牛骨一端,使与之粘连住的含水培养皿完全呈悬空状态,测量培养皿与牛骨在此种状态下完全分离所需要的时间。
如图15所示,通过在培养皿和牛骨之间均匀涂抹相同质量的水凝胶,经过在37℃条件下1h孵育后,在培养皿中注入20g去离子水,拎起牛骨一端,使与之粘连住的含水培养皿完全呈悬空状态(如图15b所示),测量培养皿与牛骨在此种状态下完全分离所需要的时间(可视为水凝胶与牛骨完全分离所需时间)。经实验测得涂抹ASP水凝胶后所需时间为50s,而A35 MO为7min。这进一步证明了A35 MO在水下粘合方面体现出更强的能力。
为了进一步探究A35 MO水下粘合的可行性,通过实验来测试其对不同材料基底(如骨骼、玻璃片)的粘合效果。分别将A35 MO均匀涂抹于断裂的骨骼、玻璃片之间,并即刻浸没于水中。结果如图25所示,证明无论对于骨骼粘合还是玻璃粘合,均能够维持至少超过8小时的水下粘合状态。此实验说明A35 MO对不同材料的均有一定的适用性。
自修复实验,主要步骤如下:
(1)取水凝胶样品于玻璃片,用锋利的刀片从水凝胶中间位置切开。
(2)将切开后的两部分水凝胶贴近,记录完全自修复的时间。
如图21(a)至(e)所示为水凝胶从完整到被切割到相互融合的过程,经过约2分钟,A35 MO即可从被破坏状态又恢复至完整状态。其自修复性质可能来源于两个方面,其一是来源于A35 MO中的物理交联作用;其二是由于A-S-Mefp3-P和A-S-Mefp5-P中多巴之间的分子间氢键作用,而研究表明甚至在水中这种作用也依然能够发生,这是由于多巴之间比多巴与水之间更容易在热力学上发生氢键的形成。
图22a所示,将A35 MO填充于一块完整的聚氨酯多孔材料。将其用刀切开后分为两半。将两部分的材料靠近,使其中的A35 MO充分接触。等待数分钟后,由于A35 MO自修复的性质,可以看到材料又归于完整,并且将其用镊子夹起一端也不会发生脱离的情况。此实验为水下粘合材料的应用带来了新的思路,即让装载着新型水下粘合材料的断裂、破损材料拥有自我修复的能力。
水下粘合材料的可注射性对于后续的应用有非常关键的作用。为了探究A35 MO是否能够具备可注射性,采用3D打印的方式来验证。主要步骤为:取水凝胶样品置于打印针筒中,选用的打印针头内径21mm,外径42mm。并将3D打印仪器参数设定为系统压强700kPa,移动速度5mm/s。
如图23a所示为用刮勺挑起A35 MO的状态,呈细丝状。图23b为A35 MO经过3D打印后形成的图案,其中六边形边长由里至外分别为3mm、5mm、7mm,圆形由里至外直径分别为3mm、5mm、7mm。通过实验证明,A35 MO成功地进行了打印实验,并且打印出来的材质均匀、规整,具备可注射性。基于实验中3D打印的成功展示,该材料有作为粘附性细胞支架的潜力。
图24为A35 MO可注射性质应用举例,如图24a所示为一块琼脂糖凝胶,其中中间部分缺失。如图24b所示,使用3D打印机将A35 MO注入缺失部分,后将三颗玻璃小球填充于缺失部分,如24c所示将材料倒置,小球紧紧地通过A35 MO粘附于琼脂糖凝胶上,不会掉落。
实施例8
A-S-Mefp3-P和A-S-Mefp5-P融合过程的探究实验:
本文的研究目标是通过同时增强材料的内在粘性与界面粘性,来提升材料整体的水下粘合能力,通过上述实施例结果分析,A35 MO增强的内在粘性,一方面依靠亮氨酸拉链的自组装性质,另一方面可能来源于A-S-Mefp3-P和A-S-Mefp5-P间的相互作用。
为了进一步验证经酪氨酸酶修饰后的A-S-Mefp3-P和A-S-Mefp5-P之间是否发生相互作用,本发明进行了一下实验:
(1)将构建完毕的A-S-Spytag-Mefp3-P和A-S-Snooptag-Mefp5-P通过前述的方式进行蛋白表达分离纯化实验,将所得A-S-Spytag-Mefp3-P与适量Mcherry-Spycatcher溶液(Mcherry-Spycatcher的序列为Mcherry的C端与Spycatcher的N端直接连接,Spycatcher氨基酸序列如SEQ ID No.16所示,Mcherry氨基酸序列如SEQ ID No.18所示)混合,A-S-Snooptag-Mefp5-P与适量GFP-Snoopcatcher(GFP-Snoopcatcher的序列为GFP的C端与Snoopcatcher的N端直接连接,Snoopcatcher氨基酸序列如SEQ ID No.17所示,GFP氨基酸序列如SEQ ID No.19所示)混合,通过前述同样方式成胶。
(2)将结合了Mcherry-Spycatcher的A-S-Spytag-Mefp3-P水凝胶(以下简称ASpy)从中间位置切开,并对结合了GFP-Snoopcatcher的A-S-Snooptag-Mefp5-水凝胶(以下简称ASnoop)进行同样操作。之后,将ASpy的一半水凝胶同Asnoop的一半水凝胶相互靠近,在荧光显微镜下观察其接触界面的现象。
结果如图20所示,20d至e显示,Aspy和Asnoop靠近的界面处发生了一些变化。从图中无箭头的椭圆框内可见,Aspy和Asnoop相互融合,体现在所具有的绿色和红色荧光融合后呈现出的黄色逐渐加深;而从图中含有箭头的椭圆框内可见,Aspy和Asnoop定点的位置间距离有所缩短。在短短10分钟内,Aspy和Asnoop即呈现出了动态的融合过程。从以上实验可以推测A-S-Mefp3-P与A-S-Mefp5-P间可能具有相互作用。后续还可在更长时间的尺度下进行同样的实验加以验证
综上所述,本发明有效克服了现有技术中的种种缺点而具高度产业利用价值。
上述实施例仅例示性说明本发明的原理及其功效,而非用于限制本发明。任何熟悉此技术的人士皆可在不违背本发明的精神及范畴下,对上述实施例进行修饰或改变。因此,举凡所属技术领域中具有通常知识者在未脱离本发明所揭示的精神与技术思想下所完成的一切等效修饰或改变,仍应由本发明的权利要求所涵盖。
序列表
<110> 上海科技大学
<120> 一种可注射自修复水下蛋白及其用途
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Gly Asn Tyr Asn Arg Tyr Gly Arg Arg Tyr Gly Gly Tyr Lys Gly Trp
20 25 30
Asn Asn Gly Trp Lys Arg Gly Arg Trp Gly Arg Lys Tyr Tyr
35 40 45
<210> 7
<211> 76
<212> PRT
<213> 人工序列Mgfp-5(Artificial Sequence)
<400> 7
Ser Ser Glu Glu Tyr Lys Gly Gly Tyr Tyr Pro Gly Asn Thr Tyr His
1 5 10 15
Tyr His Ser Gly Gly Ser Tyr His Gly Ser Gly Tyr His Gly Gly Tyr
20 25 30
Lys Gly Lys Tyr Tyr Gly Lys Ala Lys Lys Tyr Tyr Tyr Lys Tyr Lys
35 40 45
Asn Ser Gly Lys Tyr Lys Tyr Leu Lys Lys Ala Arg Lys Tyr His Arg
50 55 60
Lys Gly Tyr Lys Lys Tyr Tyr Gly Gly Gly Ser Ser
65 70 75
<210> 8
<211> 45
<212> PRT
<213> 人工序列mfp-3s(Artificial Sequence)
<400> 8
Gly Tyr Gly Tyr Asp Leu Gly Tyr Asn Ala Pro Trp Pro Tyr Asn Asn
1 5 10 15
Gly Tyr Tyr Gly Tyr Asn Gly Tyr Asn Gly Tyr His Gly Arg Tyr Gly
20 25 30
Trp Asn Lys Gly Trp Asn Asn Gly Pro Trp Gly Gly Tyr
35 40 45
<210> 9
<211> 25
<212> PRT
<213> 人工序列mfp3s-pep(Artificial Sequence)
<400> 9
Gly Tyr Asp Gly Tyr Asn Trp Pro Tyr Gly Tyr Asn Gly Tyr Arg Tyr
1 5 10 15
Gly Trp Asn Lys Gly Trp Asn Gly Tyr
20 25
<210> 10
<211> 14
<212> PRT
<213> 人工序列A1片段(Artificial Sequence)
<220>
<221> VARIANT
<222> (5)..(5)
<223> The 'Xaa' at location 5 stands for Glu, or Lys.
<220>
<221> VARIANT
<222> (7)..(7)
<223> The 'Xaa' at location 7 stands for Glu, or Lys.
<220>
<221> VARIANT
<222> (12)..(12)
<223> The 'Xaa' at location 12 stands for Glu, or Lys.
<220>
<221> VARIANT
<222> (14)..(14)
<223> The 'Xaa' at location 14 stands for Glu, or Lys.
<220>
<221> UNSURE
<222> (5)..(5)
<223> The 'Xaa' at location 5 stands for Gln, Arg, Pro, or Leu.
<220>
<221> UNSURE
<222> (7)..(7)
<223> The 'Xaa' at location 7 stands for Gln, Arg, Pro, or Leu.
<220>
<221> UNSURE
<222> (12)..(12)
<223> The 'Xaa' at location 12 stands for Gln, Arg, Pro, or Leu.
<220>
<221> UNSURE
<222> (14)..(14)
<223> The 'Xaa' at location 14 stands for Gln, Arg, Pro, or Leu.
<400> 10
Ser Gly Asp Leu Xaa Asn Xaa Val Ala Gln Leu Xaa Arg Xaa
1 5 10
<210> 11
<211> 14
<212> PRT
<213> 人工序列A1片段(Artificial Sequence)
<220>
<221> VARIANT
<222> (5)..(5)
<223> The 'Xaa' at location 5 stands for Glu, or Lys.
<220>
<221> VARIANT
<222> (7)..(7)
<223> The 'Xaa' at location 7 stands for Glu, or Lys.
<220>
<221> VARIANT
<222> (12)..(12)
<223> The 'Xaa' at location 12 stands for Glu, or Lys.
<220>
<221> VARIANT
<222> (14)..(14)
<223> The 'Xaa' at location 14 stands for Glu, or Lys.
<220>
<221> UNSURE
<222> (5)..(5)
<223> The 'Xaa' at location 5 stands for Gln, Arg, Pro, or Leu.
<220>
<221> UNSURE
<222> (7)..(7)
<223> The 'Xaa' at location 7 stands for Gln, Arg, Pro, or Leu.
<220>
<221> UNSURE
<222> (12)..(12)
<223> The 'Xaa' at location 12 stands for Gln, Arg, Pro, or Leu.
<220>
<221> UNSURE
<222> (14)..(14)
<223> The 'Xaa' at location 14 stands for Gln, Arg, Pro, or Leu.
<400> 11
Val Arg Ser Leu Xaa Asp Xaa Ala Ala Glu Leu Xaa Gln Xaa
1 5 10
<210> 12
<211> 15
<212> PRT
<213> 人工序列A1片段(Artificial Sequence)
<220>
<221> VARIANT
<222> (5)..(5)
<223> The 'Xaa' at location 5 stands for Glu, or Lys.
<220>
<221> VARIANT
<222> (7)..(7)
<223> The 'Xaa' at location 7 stands for Glu, or Lys.
<220>
<221> VARIANT
<222> (12)..(12)
<223> The 'Xaa' at location 12 stands for Glu, or Lys.
<220>
<221> VARIANT
<222> (14)..(14)
<223> The 'Xaa' at location 14 stands for Glu, or Lys.
<220>
<221> UNSURE
<222> (5)..(5)
<223> The 'Xaa' at location 5 stands for Gln, Arg, Pro, or Leu.
<220>
<221> UNSURE
<222> (7)..(7)
<223> The 'Xaa' at location 7 stands for Gln, Arg, Pro, or Leu.
<220>
<221> UNSURE
<222> (12)..(12)
<223> The 'Xaa' at location 12 stands for Gln, Arg, Pro, or Leu.
<220>
<221> UNSURE
<222> (14)..(14)
<223> The 'Xaa' at location 14 stands for Gln, Arg, Pro, or Leu.
<400> 12
Val Ser Arg Leu Xaa Asn Xaa Ile Glu Asp Leu Xaa Ala Xaa Ile
1 5 10 15
<210> 13
<211> 37
<212> PRT
<213> γ52−88KI片段(Artificial Sequence)
<400> 13
Ile Asp Phe Ile Ser Thr Tyr Ile Thr Lys Ile Asp Lys Lys Ile Gln
1 5 10 15
Ser Ile Glu Asp Ile Ile His Gln Ile Glu Asn Lys Ile Ser Glu Ile
20 25 30
Lys Gln Leu Ile Lys
35
<210> 14
<211> 13
<212> PRT
<213> 人工序列Spytag(Artificial Sequence)
<400> 14
Ala His Ile Val Met Val Asp Ala Tyr Lys Pro Thr Lys
1 5 10
<210> 15
<211> 15
<212> PRT
<213> 人工序列Snooptag(Artificial Sequence)
<400> 15
Gly Lys Leu Gly Asp Ile Glu Phe Ile Lys Val Asn Lys Gly Tyr
1 5 10 15
<210> 16
<211> 113
<212> PRT
<213> 人工序列spycatcher(Artificial Sequence)
<400> 16
Val Asp Thr Leu Ser Gly Leu Ser Ser Glu Gln Gly Gln Ser Gly Asp
1 5 10 15
Met Thr Ile Glu Glu Asp Ser Ala Thr His Ile Lys Phe Ser Lys Arg
20 25 30
Asp Glu Asp Gly Lys Glu Leu Ala Gly Ala Thr Met Glu Leu Arg Asp
35 40 45
Ser Ser Gly Lys Thr Ile Ser Thr Trp Ile Ser Asp Gly Gln Val Lys
50 55 60
Asp Phe Tyr Leu Tyr Pro Gly Lys Tyr Thr Phe Val Glu Thr Ala Ala
65 70 75 80
Pro Asp Gly Tyr Glu Val Ala Thr Ala Ile Thr Phe Thr Val Asn Glu
85 90 95
Gln Gly Gln Val Thr Val Asn Gly Lys Ala Thr Lys Gly Asp Ala His
100 105 110
Ile
<210> 17
<211> 112
<212> PRT
<213> 人工序列Snoopcatcher(Artificial Sequence)
<400> 17
Lys Pro Leu Arg Gly Ala Val Phe Ser Leu Gln Lys Gln His Pro Asp
1 5 10 15
Tyr Pro Asp Ile Tyr Gly Ala Ile Asp Gln Asn Gly Thr Tyr Gln Asn
20 25 30
Val Arg Thr Gly Glu Asp Gly Lys Leu Thr Phe Lys Asn Leu Ser Asp
35 40 45
Gly Lys Tyr Arg Leu Phe Glu Asn Ser Glu Pro Ala Gly Tyr Lys Pro
50 55 60
Val Gln Asn Lys Pro Ile Val Ala Phe Gln Ile Val Asn Gly Glu Val
65 70 75 80
Arg Asp Val Thr Ser Ile Val Pro Gln Asp Ile Pro Ala Thr Tyr Glu
85 90 95
Phe Thr Asn Gly Lys His Tyr Ile Thr Asn Glu Pro Ile Pro Pro Lys
100 105 110
<210> 18
<211> 237
<212> PRT
<213> 人工序列Mcherry(Artificial Sequence)
<400> 18
Val Ser Lys Gly Glu Glu Asp Asn Met Ala Ile Ile Lys Glu Phe Met
1 5 10 15
Arg Phe Lys Val His Met Glu Gly Ser Val Asn Gly His Glu Phe Glu
20 25 30
Ile Glu Gly Glu Gly Glu Gly Arg Pro Tyr Glu Gly Thr Gln Thr Ala
35 40 45
Lys Leu Lys Val Thr Lys Gly Gly Pro Leu Pro Phe Ala Trp Asp Ile
50 55 60
Leu Ser Pro Gln Phe Met Tyr Gly Ser Lys Ala Tyr Val Lys His Pro
65 70 75 80
Ala Asp Ile Pro Asp Tyr Leu Lys Leu Ser Phe Pro Glu Gly Phe Lys
85 90 95
Trp Glu Arg Val Met Asn Phe Glu Asp Gly Gly Val Val Thr Val Thr
100 105 110
Gln Asp Ser Ser Leu Gln Asp Gly Glu Phe Ile Tyr Lys Val Lys Leu
115 120 125
Arg Gly Thr Asn Phe Pro Ser Asp Gly Pro Val Met Gln Lys Lys Thr
130 135 140
Met Gly Trp Glu Ala Ser Ser Glu Arg Met Tyr Pro Glu Asp Gly Ala
145 150 155 160
Leu Lys Gly Glu Ile Lys Gln Arg Leu Lys Leu Lys Asp Gly Gly His
165 170 175
Tyr Asp Ala Glu Val Lys Thr Thr Tyr Lys Ala Lys Lys Pro Val Gln
180 185 190
Leu Pro Gly Ala Tyr Asn Val Asn Ile Lys Leu Asp Ile Thr Ser His
195 200 205
Asn Glu Asp Tyr Thr Ile Val Glu Gln Tyr Glu Arg Ala Glu Gly Arg
210 215 220
His Ser Thr Gly Gly Met Asp Glu Leu Tyr Lys Ala Gly
225 230 235
<210> 19
<211> 240
<212> PRT
<213> 人工序列Gfp(Artificial Sequence)
<400> 19
Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu Val
1 5 10 15
Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly Glu
20 25 30
Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile Cys
35 40 45
Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr Leu
50 55 60
Thr Tyr Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys Gln
65 70 75 80
His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu Arg
85 90 95
Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu Val
100 105 110
Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly Ile
115 120 125
Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr Asn
130 135 140
Tyr Asn Ser His Asn Val Tyr Ile Met Ala Asp Lys Gln Lys Asn Gly
145 150 155 160
Ile Lys Val Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser Val
165 170 175
Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro
180 185 190
Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Lys Leu Ser
195 200 205
Lys Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe Val
210 215 220
Thr Ala Ala Gly Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys Ala Gly
225 230 235 240
<210> 20
<211> 585
<212> DNA
<213> 人工序列A-S-P质粒(Artificial Sequence)
<400> 20
atgtctggtg acctggaaaa cgaagttgcg cagctggaac gtgaagttcg ttctctggaa 60
gacgaagcgg cggaactgga acagaaagtt tctcgtctga aaaacgaaat cgaagacctg 120
aaagcggaac catgggcagg tgccggtgca ggtccggaag gtgcaggtgc aggtgcaggt 180
ccggaaggtg caggtgcagg tgcaggtccg gagggtgccg gtgcaggtgc aggtcctgaa 240
ggtgccggtg caggtgcagg tccggaaggt gcaggtgcag gtgccggtcc tgaaggtgcc 300
ggtgcaggtg caggtcctga aggtgccggt gcaggtgccg gtcctgaggg tgcaggtgca 360
ggtgccggcc ctgaaggtgc aggtgcaggt gcaggtccgg aaggtggatc caagcttggc 420
ggtggcggta gcgctccgca gatgctgcgt gaactgcagg aaaccaacgc tgctctgcag 480
gacgttcgtg aactgctgcg tcagcaggtt aaagaaatca ccttcctgaa aaacaccgtt 540
atggaatctg acgcttctct cgagcaccac caccaccacc actga 585
<210> 21
<211> 79
<212> DNA
<213> 人工序列A-S-P-F正向引物(Artificial Sequence)
<400> 21
tacatatgtc tggtgacctg gaaaacgaag ttgcgcagct ggaacgtgaa gttcgttctc 60
tggaagacga agcggcgga 79
<210> 22
<211> 84
<212> DNA
<213> 人工序列A-S-P--R反向引物(Artificial Sequence)
<400> 22
gcccatggtt ccgctttcag gtcttcgatt tcgtttttca gacgagaaac tttctgttcc 60
agttccgccg cttcgtcttc caga 84
<210> 23
<211> 729
<212> DNA
<213> 人工序列A-S-Mefp3-P质粒(Artificial Sequence)
<400> 23
atgtctggtg acctggaaaa cgaagttgcg cagctggaac gtgaagttcg ttctctggaa 60
gacgaagcgg cggaactgga acagaaagtt tctcgtctga aaaacgaaat cgaagacctg 120
aaagcggaac catgggcagg tgccggtgca ggtccggaag gtgcaggtgc aggtgcaggt 180
ccggaaggtg caggtgcagg tgcaggtccg gagggtgccg gtgcaggtgc aggtcctgaa 240
ggtgccggtg caggtgcagg tccggaaggt gcaggtgcag gtgccggtcc tgaaggtgcc 300
ggtgcaggtg caggtcctga aggtgccggt gcaggtgccg gtcctgaggg tgcaggtgca 360
ggtgccggcc ctgaaggtgc aggtgcaggt gcaggtccgg aaggtggatc cgccgattat 420
tatggcccga attatggtcc gccgcgtcgc tacggtggcg gcaactataa ccgctacaat 480
cgctatggcc gccgctatgg tggctataaa ggctggaaca acggctggaa tcgcggtcgc 540
cgcggtaaat attggaagct tggcggtggc ggtagcgctc cgcagatgct gcgtgaactg 600
caggaaacca acgctgctct gcaggacgtt cgtgaactgc tgcgtcagca ggttaaagaa 660
atcaccttcc tgaaaaacac cgttatggaa tctgacgctt ctctcgagca ccaccaccac 720
caccactga 729
<210> 24
<211> 26
<212> DNA
<213> 人工序列A-S-Mefp3-P-F正向引物(Artificial Sequence)
<400> 24
ggatccgccg attattatgg cccgaa 26
<210> 25
<211> 25
<212> DNA
<213> 人工序列A-S-Mefp3-P-F正向引物(Artificial Sequence)
<400> 25
aagcttccaa tatttaccgc ggcga 25
<210> 26
<211> 807
<212> DNA
<213> 人工序列A-S-Mefp5-P质粒(Artificial Sequence)
<400> 26
atgtctggtg acctggaaaa cgaagttgcg cagctggaac gtgaagttcg ttctctggaa 60
gacgaagcgg cggaactgga acagaaagtt tctcgtctga aaaacgaaat cgaagacctg 120
aaagcggaac catgggcagg tgccggtgca ggtccggaag gtgcaggtgc aggtgcaggt 180
ccggaaggtg caggtgcagg tgcaggtccg gagggtgccg gtgcaggtgc aggtcctgaa 240
ggtgccggtg caggtgcagg tccggaaggt gcaggtgcag gtgccggtcc tgaaggtgcc 300
ggtgcaggtg caggtcctga aggtgccggt gcaggtgccg gtcctgaggg tgcaggtgca 360
ggtgccggcc ctgaaggtgc aggtgcaggt gcaggtccgg aaggtggatc cagcagtgaa 420
gaatataaag gcggctatta cccgggtaac gcgtaccatt accacagcgg tggtagttat 480
catggaagcg gctaccacgg tggctacaaa ggcaagtact acggtaaggc caagaaatac 540
tactacaaat acaaaaattc aggcaagtac aaatatctga agaaagcgcg caaataccac 600
cgtaaagggt ataaatacta tggtggctct tcaaagcttg gcggtggcgg tagcgctccg 660
cagatgctgc gtgaactgca ggaaaccaac gctgctctgc aggacgttcg tgaactgctg 720
cgtcagcagg ttaaagaaat caccttcctg aaaaacaccg ttatggaatc tgacgcttct 780
ctcgagcacc accaccacca ccactga 807
<210> 27
<211> 26
<212> DNA
<213> 人工序列A-S-Mefp5-P-F正向引物(Artificial Sequence)
<400> 27
ggatccagca gtgaagaata taaagg 26
<210> 28
<211> 28
<212> DNA
<213> 人工序列A-S-Mefp5-P-R反向引物(Artificial Sequence)
<400> 28
aagctttgaa gagccaccat agtattta 28
<210> 29
<211> 768
<212> DNA
<213> 人工序列A-S-Spytag-Mefp3-P质粒(Artificial Sequence)
<400> 29
atgtctggtg acctggaaaa cgaagttgcg cagctggaac gtgaagttcg ttctctggaa 60
gacgaagcgg cggaactgga acagaaagtt tctcgtctga aaaacgaaat cgaagacctg 120
aaagcggaac catgggcagg tgccggtgca ggtccggaag gtgcaggtgc aggtgcaggt 180
ccggaaggtg caggtgcagg tgcaggtccg gagggtgccg gtgcaggtgc aggtcctgaa 240
ggtgccggtg caggtgcagg tccggaaggt gcaggtgcag gtgccggtcc tgaaggtgcc 300
ggtgcaggtg caggtcctga aggtgccggt gcaggtgccg gtcctgaggg tgcaggtgca 360
ggtgccggcc ctgaaggtgc aggtgcaggt gcaggtccgg aaggtggatc cgcgcacatc 420
gttatggtcg atgcatataa acccaccaaa gccgattatt atggcccgaa ttatggtccg 480
ccgcgtcgct acggtggcgg caactataac cgctacaatc gctatggccg ccgctatggt 540
ggctataaag gctggaacaa cggctggaat cgcggtcgcc gcggtaaata ttggaagctt 600
ggcggtggcg gtagcgctcc gcagatgctg cgtgaactgc aggaaaccaa cgctgctctg 660
caggacgttc gtgaactgct gcgtcagcag gttaaagaaa tcaccttcct gaaaaacacc 720
gttatggaat ctgacgcttc tctcgagcac caccaccacc accactga 768
<210> 30
<211> 65
<212> DNA
<213> 人工序列A-S-Spytag-Mefp3-P-F正向引物(Artificial Sequence)
<400> 30
ggatccgcgc acatcgttat ggtcgatgca tataaaccca ccaaagccga ttattatggc 60
ccgaa 65
<210> 31
<211> 28
<212> DNA
<213> 人工序列A-S-Spytag-Mefp3-P-R反向引物(Artificial Sequence)
<400> 31
aagcttccaa tatttaccgc ggcgaccg 28
<210> 32
<211> 852
<212> DNA
<213> 人工序列A-S-Snooptag-Mefp5-P质粒(Artificial Sequence)
<400> 32
atgtctggtg acctggaaaa cgaagttgcg cagctggaac gtgaagttcg ttctctggaa 60
gacgaagcgg cggaactgga acagaaagtt tctcgtctga aaaacgaaat cgaagacctg 120
aaagcggaac catgggcagg tgccggtgca ggtccggaag gtgcaggtgc aggtgcaggt 180
ccggaaggtg caggtgcagg tgcaggtccg gagggtgccg gtgcaggtgc aggtcctgaa 240
ggtgccggtg caggtgcagg tccggaaggt gcaggtgcag gtgccggtcc tgaaggtgcc 300
ggtgcaggtg caggtcctga aggtgccggt gcaggtgccg gtcctgaggg tgcaggtgca 360
ggtgccggcc ctgaaggtgc aggtgcaggt gcaggtccgg aaggtggatc cggaaaactg 420
ggggacatcg aattcatcaa agtaaacaaa ggttacagca gtgaagaata taaaggcggc 480
tattacccgg gtaacgcgta ccattaccac agcggtggta gttatcatgg aagcggctac 540
cacggtggct acaaaggcaa gtactacggt aaggccaaga aatactacta caaatacaaa 600
aattcaggca agtacaaata tctgaagaaa gcgcgcaaat accaccgtaa agggtataaa 660
tactatggtg gctcttcaaa gcttggcggt ggcggtagcg ctccgcagat gctgcgtgaa 720
ctgcaggaaa ccaacgctgc tctgcaggac gttcgtgaac tgctgcgtca gcaggttaaa 780
gaaatcacct tcctgaaaaa caccgttatg gaatctgacg cttctctcga gcaccaccac 840
caccaccact ga 852
<210> 33
<211> 71
<212> DNA
<213> 人工序列A-S-Snooptag-Mefp5-P-F正向引物(Artificial Sequence)
<400> 33
ggatccggaa aactggggga catcgaattc atcaaagtaa acaaaggtta cagcagtgaa 60
gaatataaag g 71
<210> 34
<211> 29
<212> DNA
<213> 人工序列A-S-Snooptag-Mefp5-P-R反向引物(Artificial Sequence)
<400> 34
aagctttgaa gagccaccat agtatttat 29
Claims (12)
1.一种融合蛋白,所述融合蛋白包括贻贝足丝蛋白片段和卷曲螺旋结构片段。
2.如权利要求1所述的融合蛋白,其特征在于,还包括如下技术特征中的一个或多个:
A1)所述贻贝足丝蛋白选自紫贻贝足丝蛋白或地中海贻贝足丝蛋白;
A2)所述贻贝足丝蛋白选自mefp-3、mefp-5、Mgfp-3、Mgfp-5、mfp-3s或mfp3s-pep;,
A3)所述贻贝足丝蛋白片段为:a)氨基酸序列如SEQ ID No.4-9所示的多肽片段;
或,b)氨基酸序列与SEQ ID NO.4-9具有80%以上同源性、且具有a)限定的多肽片段的功能的多肽片段;
A4)所述贻贝足丝蛋白来源于贻贝;
A5)所述卷曲螺旋结构片段选自亮氨酸拉链片段、A1片段、ZE/ZR片段、GCN4片段、γ52- 88KI片段、[(AG)3PEG]10片段、PCxP片段、NLP片段;
A6)所述亮氨酸拉链片段包括A片段、S片段和P片段;
所述A片段为:c)氨基酸序列如SEQ ID No.1所示的多肽片段;或,d)氨基酸序列与SEQID NO.1具有80%以上同源性、且具有c)限定的多肽片段的功能的多肽片段;
所述S片段为:e)氨基酸序列如SEQ ID No.2所示的多肽片段;或,f)氨基酸序列与SEQID NO.2具有80%以上同源性、且具有e)限定的多肽片段的功能的多肽片段;
所述P片段为:g)氨基酸序列如SEQ ID No.3所示的多肽片段;或,h)氨基酸序列与SEQID NO.3具有80%以上同源性、且具有g)限定的多肽片段的功能的多肽片段;
A6)所述融合蛋白还包括功能性蛋白片段。
3.如权利要求2所述的融合蛋白,其特征在于,还包括如下技术特征中的一个或多个:
B1)所述亮氨酸拉链片段自N端至C端依次包括A片段、S片段和P片段;
B2)所述贻贝足丝蛋白片段被插入亮氨酸拉链片段中;
B3)所述融合蛋白自N端至C端依次包括A片段、S片段、贻贝足丝蛋白片段和P片段;
B4)所述融合蛋白经酪氨酸酶修饰;
B5)所述功能性蛋白片段选自粘性蛋白片段、细胞粘附多肽片段、酶蛋白片段、生长因子片段、抗菌活性多肽片段、凝血因子片段、生物相容性多肽片段、抗生肽片段、监测多肽片段中的一种或多种的组合;
B6)所述融合蛋白自N端至C端依次包括A片段、S片段、功能性蛋白片段、贻贝足丝蛋白片段和P片段。
4.一种分离的多核苷酸,编码如权利要求1-3任一权利要求所述的融合蛋白。
5.一种构建体,所述构建体含有权利要求6所述的分离的多核苷酸。
6.一种表达系统,所述表达系统含有如权利要求7所述的构建体或基因组中整合有外源的如权利要求6所述的多核苷酸。
7.如权利要求1-3任一权利要求所述的融合蛋白的制备方法,包括:在适合表达所述融合蛋白的条件下,培养如权利要求7所述的表达系统。
8.如权利要求7所述的制备方法,其特征在于,将表达所得的融合蛋白进行酪氨酸酶修饰。
9.一种自组装胶体溶液,所述溶液包括如权利要求1-3任一权利要求所述的融合蛋白。
10.一种胶体,所述胶体包括如权利要求1-3任一权利要求所述的融合蛋白。
11.如权利要求10所述的胶体,其特征在于,还包括如下技术特征中的一个或多个:
C1)所述胶体为水凝胶;
C2)胶体中融合蛋白的含量为70mg/mL~250mg/mL;
C3)所述胶体由所述融合蛋白自组装获得;
C4)所述胶体由权利要求9所述的自组装胶体溶液制备获得;
C5)所述胶体中,融合蛋白中的贻贝足丝蛋白选自mefp-3和mefp-5的组合。
12.如权利要求1-3任一权利要求所述的融合蛋白、如权利要求9所述的自组装胶体溶液、如权利要求10-11任一权利要求所述的胶体在水下粘合剂制备领域的用途。
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Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111505175A (zh) * | 2019-01-30 | 2020-08-07 | 上海科技大学 | 一种质谱用交联剂及其制备与应用 |
CN112625105A (zh) * | 2021-01-12 | 2021-04-09 | 厦门绥之科技有限公司 | 一种贻贝黏蛋白的生物制备方法 |
CN117247441A (zh) * | 2023-11-16 | 2023-12-19 | 北京未名拾光生物技术有限公司 | 重组贻贝粘蛋白及其表达体系和应用 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20160220727A1 (en) * | 2013-11-13 | 2016-08-04 | Massachusetts Institute Of Technology | Self-assembling underwater adhesives |
CN107446052A (zh) * | 2017-06-09 | 2017-12-08 | 上海科技大学 | 多功能复合超分子纳米纤维自组装体系及其应用 |
-
2018
- 2018-07-13 CN CN201810776842.4A patent/CN108948208A/zh active Pending
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20160220727A1 (en) * | 2013-11-13 | 2016-08-04 | Massachusetts Institute Of Technology | Self-assembling underwater adhesives |
CN107446052A (zh) * | 2017-06-09 | 2017-12-08 | 上海科技大学 | 多功能复合超分子纳米纤维自组装体系及其应用 |
Non-Patent Citations (2)
Title |
---|
WEI SHEN等: "Tuning the erosion rate of artificial protein hydrogels through control of network topology", 《NAT MATER》 * |
赵田鑫等: "合成生物学技术在材料科学中的应用", 《生物工程学报》 * |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111505175A (zh) * | 2019-01-30 | 2020-08-07 | 上海科技大学 | 一种质谱用交联剂及其制备与应用 |
CN111505175B (zh) * | 2019-01-30 | 2022-07-05 | 上海科技大学 | 一种质谱用交联剂及其制备与应用 |
CN112625105A (zh) * | 2021-01-12 | 2021-04-09 | 厦门绥之科技有限公司 | 一种贻贝黏蛋白的生物制备方法 |
CN117247441A (zh) * | 2023-11-16 | 2023-12-19 | 北京未名拾光生物技术有限公司 | 重组贻贝粘蛋白及其表达体系和应用 |
CN117247441B (zh) * | 2023-11-16 | 2024-02-02 | 北京未名拾光生物技术有限公司 | 重组贻贝粘蛋白及其表达体系和应用 |
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