CN105784656A - 一种检测活细胞内RhoGDIα蛋白活性的生物探针 - Google Patents
一种检测活细胞内RhoGDIα蛋白活性的生物探针 Download PDFInfo
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Abstract
本发明属于细胞生物学及分子生物学技术领域,涉及一种根据FRET基本原理设计制备的、可检测活细胞内RhoGDIα蛋白活性的生物探针。该探针为融合的蛋白质结构,包括FRET荧光蛋白对ECFP与Ypet,RhoGDIα全长序列,RhoGDIα的底物结构域switch II及中间的linker序列五个部分。本发明的融合质粒转入活细胞后,可由细胞自行表达出探针的融合蛋白结构,从而在细胞内发挥探针功能,具有特异性高、成本低、对细胞无副作用等特点。
Description
技术领域
本发明属于细胞生物学及分子生物学技术领域,是基于荧光共振能量转移(fluorescenceresonanceenergytransfer,FRET)原理和亚克隆技术进行设计制备,通过荧光蛋白颜色与强度变化检测活细胞内RhoGDIα(RhoGDPdissociationinhibitorα)蛋白活性变化的生物探针。
背景技术
RhoGDIα是Rho家族成员的负性调控蛋白,可通过抑制Rho家族不同成员的活性参与细胞迁移过程,对癌细胞侵袭具有至关重要的作用。目前一般采用基于蛋白抗原抗体结合的免疫组织化学反应、WesternBlotting检测、或基于核酸水平的原位PCR反应等手段检测细胞内RhoGDIα蛋白,这些手段都存在一定的缺陷:1.必须对细胞进行破碎或固定等预处理,破坏了细胞的正常生理状态;2.不能在单细胞或亚细胞水平上动态观测;3.只能检测蛋白的量多少,不能检测蛋白的活性高低;4.成本高。因此,目前没有一种切实有效的方法能对活细胞内RhoGDIα活性进行检测。针对上述缺点,本发明提出一种基于FRET技术的生物探针,可以检测活细胞中RhoGDIα活性变化,同时具备以下优点:1.亚克隆技术大大降低了探针制备周期及成本;2.可在单细胞及亚细胞水平上进行观测;3.可以对同一细胞进行实时动态检测;4.在活细胞上观察,不伤害细胞,能反映细胞内的真实状态。
发明内容
本发明提供一种可检测活细胞中RhoGDIα活性的新型生物探针,设计中将FRET技术与亚克隆技术结合,利用RhoGDIα活性改变能与其底物结合而改变探针空间结构,从而引起FRET效率发生变化的特点,使其能实时检测出单一活细胞中RhoGDIα活性及空间分布的动态变化。
本发明的技术方案如下:
如附图所示,该RhoGDIα-FRET探针为蛋白质,结构共包括FRET荧光蛋白对ECFP(1-1)与Ypet(1-5),RhoGDIα全长蛋白(1-4),Rho家族共同底物序列switchII(1-2)及连接linker序列(1-3)。在设计及制备过程中,采用亚克隆技术对FRET荧光蛋白对ECFP(1-1)、RhoGDIα的底物结构域switchII(1-2)、linker序列(1-3)、RhoGDIα全长序列(1-4)和Ypet(1-5)五个部分在DNA水平上进行连接,并包装入商品化的pcDNA3.1质粒,从而构建出含RhoGDIα-FRET探针DNA序列的融合质粒,该质粒用常规方法转染活细胞,即可在细胞内自行表达出探针蛋白。通过FRET显微镜观察荧光,从而发挥检测功能。
其中,RhoGDIα全长氨基酸序列如下:
MAEQEPTAEQLAQIAAENEEDEHSVNYKPPAQKSIQEIQELDKDDESLRKYKEALLGRVAVSADPNVPNVIVTRLTLVCSTAPGPLELDLTGDLESFKKQSFVLKEGVEYRIKISFRVNREIVSGMKYIQHTYRKGVKIDKTDYMVGSYGPRAEEYEFLTPMEEAPKGMLARGSYNIKSRFTDDDKTDHLSWEWNLTIKKEWKD
其对应的DNA序列为:
ATGGCAGAACAGGAACCCACTGCTGAGCAGCTCGCTCAGATAGCTGCAGAGAATGAGGAAGACGAGCACTCTGTGAACTACAAGCCTCCAGCCCAGAAGAGCATCCAGGAGATCCAGGAACTGGACAAGGATGATGAAAGCCTTCGAAAGTACAAGGAGGCCCTGCTGGGCCGAGTAGCTGTCTCTGCAGACCCCAATGTCCCCAACGTCATTGTGACCCGCTTGACCTTGGTGTGCAGCACTGCCCCAGGCCCTCTGGAACTGGACCTGACAGGTGATCTGGAGAGCTTCAAGAAACAGTCATTTGTGTTGAAGGAAGGTGTGGAGTACCGGATAAAAATCTCTTTCCGGGTGAACAGAGAGATCGTGTCAGGCATGAAGTACATCCAGCACACATACAGGAAAGGGGTCAAAATTGACAAGACTGACTACATGGTCGGGAGCTATGGGCCAAGGGCTGAGGAGTATGAGTTCCTGACACCCATGGAAGAGGCCCCCAAAGGCATGCTTGCTCGGGGCAGTTACAACATCAAGTCCCGCTTCACAGATGATGACAAAACTGACCACCTGTCCTGGGAGTGGAATCTCACCATCAAAAAGGAATGGAAGGACTGA
Rho家族共有底物序列SwitchII氨基酸序列全长:
DTAGQEDYDRLRPLS
对应的DNA序列为:
GATACTGCAGGGCAAGAGGATTATGACAGACTACGACCGTTATCC
Linker氨基酸序列为:GGSGGT
对应的DNA序列为:GGTGGCTCTGGCGGTACT。
该生物探针的融合质粒转入活细胞内,由细胞自行表达出探针的融合蛋白结构,在活细胞内通过FRET荧光显微镜动态检测RhoGDIα蛋白活性及分布情况变化。
本发明的有益效果:该探针具有特异性高、成本低、对细胞无副作用等特点,实现了在单一活细胞中对RhoGDIα活性变化及空间分布情况的实时观测,是一种廉价高效的FRET生物探针,为科学研究或者临床检测细胞内RhoGDIα活性状态提供一种工具。
附图说明
图1是本发明的RhoGDIα-FRET探针结构示意图。
图2是本发明的RhoGDIα-FRET探针工作原理图。
图中:1-1FRET荧光蛋白对ECFP;1-2RhoGDIα的底物结构域switchII;1-3linker序列;1-4RhoGDIα全长序列;1-5Ypet。
具体实施方式
以下结合附图和技术方案,进一步说明本发明的具体实施方式。
本探针可在真核细胞中自行表达,能准确反映RhoGDIα活性及空间分布的变化情况且不影响细胞自身功能。具体实施方法如下:对真核细胞转染RhoGDIα-FRET探针的融合质粒,细胞即自行表达RhoGDIα-FRET探针的融合蛋白。当该探针上的RhoGDIα蛋白(1-4)具备活性时,可与探针上的底物switchII(1-2)相互结合,整个探针发生折叠改变探针的空间结构,探针上的ECFP(1-1)与Ypet(1-5)之间的距离改变,就会影响两个荧光蛋白之间的能量转移效率。在FRET显微镜下对转入RhoGDIα-FRET探针的活细胞拍照,以波长420nm的激发光进行激发,同时在485nm、535nm波长上采集荧光图像,通过485nm/535nm荧光强度比例分析能量转移的效率变化,即可获得活细胞中RhoGDIα活性及空间分布情况。
Claims (2)
1.一种检测活细胞内RhoGDIα蛋白活性的生物探针,其特征在于,该生物探针为融合的蛋白质结构,包括FRET荧光蛋白对ECFP(1-1)与Ypet(1-5),RhoGDIα全长序列(1-4),RhoGDIα的底物结构域switchII(1-2)及linker序列(1-3)五个部分;FRET荧光蛋白对ECFP(1-1)、RhoGDIα的底物结构域switchII(1-2)、linker序列(1-3)、RhoGDIα全长序列(1-4)和Ypet(1-5)依次连接;
其中,RhoGDIα全长序列的氨基酸序列如下:
MAEQEPTAEQLAQIAAENEEDEHSVNYKPPAQKSIQEIQELDKDDESLRKYKEALLGRVAVSADPNVPNVIVTRLTLVCSTAPGPLELDLTGDLESFKKQSFVLKEGVEYRIKISFRVNREIVSGMKYIQHTYRKGVKIDKTDYMVGSYGPRAEEYEFLTPMEEAPKGMLARGSYNIKSRFTDDDKTDHLSWEWNLTIKKEWKD
其对应的DNA序列为:
ATGGCAGAACAGGAACCCACTGCTGAGCAGCTCGCTCAGATAGCTGCAGAGAATGAGGAAGACGAGCACTCTGTGAACTACAAGCCTCCAGCCCAGAAGAGCATCCAGGAGATCCAGGAACTGGACAAGGATGATGAAAGCCTTCGAAAGTACAAGGAGGCCCTGCTGGGCCGAGTAGCTGTCTCTGCAGACCCCAATGTCCCCAACGTCATTGTGACCCGCTTGACCTTGGTGTGCAGCACTGCCCCAGGCCCTCTGGAACTGGACCTGACAGGTGATCTGGAGAGCTTCAAGAAACAGTCATTTGTGTTGAAGGAAGGTGTGGAGTACCGGATAAAAATCTCTTTCCGGGTGAACAGAGAGATCGTGTCAGGCATGAAGTACATCCAGCACACATACAGGAAAGGGGTCAAAATTGACAAGACTGACTACATGGTCGGGAGCTATGGGCCAAGGGCTGAGGAGTATGAGTTCCTGACACCCATGGAAGAGGCCCCCAAAGGCATGCTTGCTCGGGGCAGTTACAACATCAAGTCCCGCTTCACAGATGATGACAAAACTGACCACCTGTCCTGGGAGTGGAATCTCACCATCAAAAAGGAATGGAAGGACTGA;
RhoGDIα的底物结构域switchII的氨基酸序列全长:
DTAGQEDYDRLRPLS
对应的DNA序列为:
GATACTGCAGGGCAAGAGGATTATGACAGACTACGACCGTTATCC;
Linker氨基酸序列为:GGSGGT
对应的DNA序列为:GGTGGCTCTGGCGGTACT。
2.根据权利要求1所述的生物探针的应用,其特征在于,该生物探针的融合质粒转入活细胞内,由细胞自行表达出探针的融合蛋白结构,在活细胞内通过FRET荧光显微镜动态检测RhoGDIα蛋白活性及分布情况变化。
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Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106399338A (zh) * | 2016-08-29 | 2017-02-15 | 大连理工大学 | 一种检测活细胞细胞膜表面张力变化的生物探针 |
CN110616255A (zh) * | 2019-10-14 | 2019-12-27 | 康妍葆(北京)干细胞科技有限公司 | Rna水平鉴别牙髓干细胞与牙龈干细胞的引物组、试剂盒及鉴定方法 |
CN112661859A (zh) * | 2020-12-23 | 2021-04-16 | 大连理工大学 | 一种基于fret的活细胞内pim蛋白活性检测生物探针 |
CN115947866A (zh) * | 2022-09-28 | 2023-04-11 | 大连理工大学 | 一种基于FRET的活细胞内Paxillin蛋白活性检测生物探针及其重组质粒 |
Citations (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2000279054A (ja) * | 1999-03-30 | 2000-10-10 | Japan Science & Technology Corp | RhoGDI遺伝子欠損動物 |
WO2002022565A1 (en) * | 2000-09-15 | 2002-03-21 | Iconix Pharmaceuticals, Inc. | Inhibitors of rho c |
GB2375538A (en) * | 2001-02-15 | 2002-11-20 | Glaxo Group Ltd | Polypeptide constructs for FRET analysis |
WO2003043577A2 (en) * | 2001-11-19 | 2003-05-30 | Iconix Pharmaceuticals, Inc. | Modulators of rho c activity |
US20040053328A1 (en) * | 2000-08-14 | 2004-03-18 | Michiyuki Matsuda | Monitoring proteins for the activities of low-molecular- weight gtp-binding proteins |
CN1685234A (zh) * | 2002-09-27 | 2005-10-19 | 阿勒根公司 | 基于细胞的荧光共振能量传递(fret)检测梭菌毒素 |
US20060029946A1 (en) * | 2004-03-12 | 2006-02-09 | Hahn Klaus M | Live cell biosensors |
US20060265764A1 (en) * | 2003-04-24 | 2006-11-23 | Yoshio Umezawa | Probe for detecting and quantifying lipid second messenger and method of detecting and quantifying lipid second messenger using the same |
US7834148B2 (en) * | 2006-05-23 | 2010-11-16 | Stanford University | Protein phosphorylation imaging systems, methods of making phosphorylation imaging systems, and methods of use thereof |
US20110071045A1 (en) * | 2009-09-22 | 2011-03-24 | William Patterson | Novel method for the selection of specific affinity binders by homogeneous noncompetitive assay |
CN103228669A (zh) * | 2010-09-27 | 2013-07-31 | 国立大学法人京都大学 | 基于荧光共振能量转移的原理的单分子型fret生物传感器接头 |
CN104762385A (zh) * | 2015-04-02 | 2015-07-08 | 王维山 | 一种检测uPA功能的FRET生物传感器及其构建方法与应用 |
-
2016
- 2016-03-16 CN CN201610150171.1A patent/CN105784656B/zh active Active
Patent Citations (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2000279054A (ja) * | 1999-03-30 | 2000-10-10 | Japan Science & Technology Corp | RhoGDI遺伝子欠損動物 |
US20040053328A1 (en) * | 2000-08-14 | 2004-03-18 | Michiyuki Matsuda | Monitoring proteins for the activities of low-molecular- weight gtp-binding proteins |
WO2002022565A1 (en) * | 2000-09-15 | 2002-03-21 | Iconix Pharmaceuticals, Inc. | Inhibitors of rho c |
GB2375538A (en) * | 2001-02-15 | 2002-11-20 | Glaxo Group Ltd | Polypeptide constructs for FRET analysis |
WO2003043577A2 (en) * | 2001-11-19 | 2003-05-30 | Iconix Pharmaceuticals, Inc. | Modulators of rho c activity |
CN1685234A (zh) * | 2002-09-27 | 2005-10-19 | 阿勒根公司 | 基于细胞的荧光共振能量传递(fret)检测梭菌毒素 |
US20060265764A1 (en) * | 2003-04-24 | 2006-11-23 | Yoshio Umezawa | Probe for detecting and quantifying lipid second messenger and method of detecting and quantifying lipid second messenger using the same |
US20060029946A1 (en) * | 2004-03-12 | 2006-02-09 | Hahn Klaus M | Live cell biosensors |
US7834148B2 (en) * | 2006-05-23 | 2010-11-16 | Stanford University | Protein phosphorylation imaging systems, methods of making phosphorylation imaging systems, and methods of use thereof |
US20110071045A1 (en) * | 2009-09-22 | 2011-03-24 | William Patterson | Novel method for the selection of specific affinity binders by homogeneous noncompetitive assay |
CN103228669A (zh) * | 2010-09-27 | 2013-07-31 | 国立大学法人京都大学 | 基于荧光共振能量转移的原理的单分子型fret生物传感器接头 |
CN104762385A (zh) * | 2015-04-02 | 2015-07-08 | 王维山 | 一种检测uPA功能的FRET生物传感器及其构建方法与应用 |
Non-Patent Citations (2)
Title |
---|
KIM ET AL.: "Substrate Rigidity Regulates Ca2+ Oscillation Via RhoA Pathway in Stem Cells", 《JOURNAL OF CELLULAR PHYSIOLOGY》 * |
OLIVIER PERTZ ET AL.: "Spatiotemporal dynamics of RhoA activity in migrating cells", 《NATURE》 * |
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106399338A (zh) * | 2016-08-29 | 2017-02-15 | 大连理工大学 | 一种检测活细胞细胞膜表面张力变化的生物探针 |
CN106399338B (zh) * | 2016-08-29 | 2019-05-14 | 大连理工大学 | 一种检测活细胞细胞膜表面张力变化的生物探针 |
CN110616255A (zh) * | 2019-10-14 | 2019-12-27 | 康妍葆(北京)干细胞科技有限公司 | Rna水平鉴别牙髓干细胞与牙龈干细胞的引物组、试剂盒及鉴定方法 |
CN110616255B (zh) * | 2019-10-14 | 2023-01-24 | 康妍葆(北京)干细胞科技有限公司 | Rna水平鉴别牙髓干细胞与牙龈干细胞的引物组、试剂盒及鉴定方法 |
CN112661859A (zh) * | 2020-12-23 | 2021-04-16 | 大连理工大学 | 一种基于fret的活细胞内pim蛋白活性检测生物探针 |
CN115947866A (zh) * | 2022-09-28 | 2023-04-11 | 大连理工大学 | 一种基于FRET的活细胞内Paxillin蛋白活性检测生物探针及其重组质粒 |
CN115947866B (zh) * | 2022-09-28 | 2024-04-19 | 大连理工大学 | 一种基于FRET的活细胞内Paxillin蛋白活性检测生物探针及其重组质粒 |
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