CN103483415A - Method for marking nanoribbon protein by quantum dots - Google Patents
Method for marking nanoribbon protein by quantum dots Download PDFInfo
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- CN103483415A CN103483415A CN201310428686.XA CN201310428686A CN103483415A CN 103483415 A CN103483415 A CN 103483415A CN 201310428686 A CN201310428686 A CN 201310428686A CN 103483415 A CN103483415 A CN 103483415A
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Abstract
The invention discloses a method for marking nanoribbon protein by quantum dots and belongs to the technical field of nano-biology. According to the invention, a mixed nanoribbon protein which contains histidine and curls irregularly is designed, and complementary polypeptides are added, so that the irregularly curling sequence of the nanoribbon protein is converted to be rigid and spiral curling and the histidine is enabled to be on one side of the spiral curling, and then the spiral curlingprotein is mixed with the quantum dots, so as to obtain a quantum dot marked composite. According to the method provided by the invention, the stability of the combining of the protein and the quantum dots can be improved. The quantum dots and the protein can be used as nanometer fluorescence probes extensively used as biological markers.
Description
Technical field
The present invention relates to the nanometer biotechnology field, be specifically related to a kind of method of quantum dot-labeled nano belt albumen.
Background technology
Quantum dot (QDs) is a kind of zero dimension semiconductor nanocrystal, almost spherical, and diameter 1～12nm, can be scattered in water or organic solvent and form glue.With traditional organic dye, compare, QDs has more excellent spectral quality, and as exciting light spectrum width and continuous distribution, and emmission spectrum is symmetric and width is narrow, color is adjustable, and photochemical stability is high, be difficult for photodissociation (Marcel BJ, Mario M, Peter G, et al.Science1998,281,2013-2016).These superior performances all are widely applied the aspect such as cell imaging and biological specific markers QDs in vivo, and it also will become very important a kind of probe tool in the molecular cell imaging research day by day.
At present, most widely used in biomarker is the synthetic fat-soluble CdSe/ZnS quantum dot of metal organic solvent method.Therefore, fat-soluble QDs must be changed into to water miscible QDs and just can carry out biomarker.And fat-soluble QDs is changed into to the most frequently used method of water miscible QDs, be first to use sulfydryl small molecules (as Thiovanic acid, thiohydracrylic acid, gsh, dimercaptosuccinic acid etc.) to replace the part on fat-soluble QDs surface, then by coupling agent and biomolecules coupling; The method of the QDs mark biomolecules that another kind is commonly used is and contains histidine-tagged polypeptide or protein binding.The affine coordinative role of metal of the Zn atom on QDs surface and polyhistidine residue, the proteins and peptides that contains histidine residues can directly be received the Zn atom on QDs surface, this method has good stability, becomes gradually a kind of method commonly used in biomarker.(the Sapsford K.E. such as Sapsford, Pons T., Medintz I.L., et al.J.Phys.Chem.C2007,111,11528-11538) systematically studied containing the interaction between Histidine polypeptide and QDs and binding constant etc., the association rate of 6 Histidine sequences and QDs is approximately 100 seconds, K
d -1be about 1nM.Up to the present, someone studies the variation of protein structure to quantum dot-labeled impact not yet.Thereby, we have designed a kind of nano belt albumen of the random coil that contains Histidine, and utilize complementary polypeptide to mediate it, make the nano belt protein structure change the spiral curling of rigidity into by random coil, nano belt albumen after conversion is more easily assembled with quantum dot, and this work is conducive to design protein ligands for nanoparticle.
Summary of the invention
The technical problem to be solved in the present invention is: for the variation of studying protein structure on quantum dot-labeled impact, and the problem of QDs labelled protein poor stability.
For solving the problems of the technologies described above, the invention provides a kind of method of quantum dot-labeled nano belt albumen, design a kind of fusion nano belt albumen of the random coil that contains Histidine, and add complementary polypeptide, make the random coil sequence of nano belt albumen be converted into the spiral curling of rigidity, make Histidine all in a spiral curling side, then spiral curling albumen is mixed with quantum dot, obtain quantum dot-labeled mixture.
Merging nano belt albumen is to add one section peptide sequence by an albumen to be marked to form again, and for example, this section peptide sequence is to be connected by proline(Pro) by 5 Pep1.Now 5 Pep1 sequences of this section are random, after adding complementary polypeptide, after adding the complementary polypeptide Pep2 of 5 times of molecular weight, this section sequence from random become spiral curling, just as a band, i.e. so-called nano belt albumen.The effect connected by proline(Pro) is the angle that a rotation can be provided, and makes its final nano belt that forms.
The sequence of described pep1 is HK VAQLKHE NQALEHE VASLEHK VSAL(SEQ ID NO.1).
Described pep2 is KEVQALEEK NAQLKEK VSALKKE VASLE(SEQ ID NO.2).
Due to form spiral curling after, Histidine, in a spiral curling side, can be combined with quantum dot simultaneously, thereby improves the stability of combination.
Nano belt albumen after conversion is looped around the quantum dot surface after quantum dot is combined.
Quantum dot of the present invention is the quantum dot that contains Zn, as CdSe/ZnS, CdTe/ZnS, CdSe/ZnSe or CdTe/ZnSe.
After adopting above-mentioned technical scheme, the beneficial effect that the present invention obtains is, the method of a kind of quantum dot-labeled nano belt albumen provided by the invention, working method is simple, repeatable high, can change protein structure it easily is combined with quantum dot, be conducive to for nanoparticle designs the novel protein part, further expand the application in biology as namo fluorescence probe of quantum dot-albumen.
The accompanying drawing explanation
Below in conjunction with drawings and Examples, the present invention is further described.
The system specialization of Fig. 1 nano belt albumen; (a) the nano belt albumen of random coil; (b) add pep2 (KEVQALEEK NAQLKEK VSALKKE VASLE) to form spiral curling; (c) spiral curling nano belt albumen and QD assembling; (d) helix wheel of spiral coiled structure is described and is shown that Histidine all is arranged in spiral curling one side.
The spiral curling nano belt albumen of Fig. 2 is easier to the quantum dot combination; A, without complementary polypeptide; B, have complementary polypeptide.
Embodiment
Embodiment
The present invention will be described further with regard to following examples, but will be appreciated that, these embodiment are the use for illustrating only, and should not be interpreted as restriction of the invention process.
Embodiment 1
The fusion nano belt albumen of design, be comprised of two portions: (1) mcherry fluorescin; (2) 5 random coil peptide sequence pep1 connect by proline(Pro), and each pep1 comprises 4 Histidines.When adding polypeptide (pep2, KEVQALEEK NAQLKEK VSALKKE VASLE) after, the random coil sequence of nano belt albumen is converted to spiral curling, make Histidine all in a spiral curling side, thereby the albumen after conversion is easier and quantum dot is assembled, and encircles in the quantum dot surface.
The transformation of nano belt albumen configuration
Design pep1 is HK VAQLKHE NQALEHE VASLEHK VSAL, and 5 pep1 are connected by proline(Pro), with the mcherry fluorescin, forms and merges nano belt albumen.
The pep2 of 5eq (KEVQALEEK NAQLKEK VSALKKE VASLE) is mixed with nano belt albumen, and nano belt albumen changes rigidity spiral curling (Fig. 1) into by original random coil.
The CdSe/ZnS quantum dot that quantum dot used is emission wavelength 610nm.
Histidine in the nano belt albumen of random coil is scattered and arranges at random, and the Histidine in spiral curling nano belt albumen all is arranged in spiral curling one side (Fig. 1 (d)).
Spiral curling nano belt albumen and quantum point coupling, the albumen after coupling just is surrounded on quantum dot surface (Fig. 1 (c)).And, after spiral curling nano belt albumen and quantum point coupling, at 610nm, FRET signal (Fig. 2, curve b) clearly appears, the nano belt albumen that therefore confirms spiral curling nano belt protein ratio random coil more easily is combined (Fig. 2) with quantum dot.
Embodiment 2
The CdTe/ZnS quantum dot that quantum dot used is emission wavelength 650nm, other steps are with embodiment 1.
Embodiment 3
Fusion rotein used is the GFP fluorescin, and other steps are with embodiment 1.
By above-mentioned three embodiment, can find out, the present invention can realize the quick combination of quantum dot and nano belt albumen, and stability is high, easy to operate.
The above-mentioned foundation desirable embodiment of the present invention of take is enlightenment, and by above-mentioned description, the relevant staff can, in the scope that does not depart from this invention technological thought, carry out various change and modification fully.The technical scope of this invention is not limited to the content on specification sheets, must determine its technical scope according to the claim scope.
Claims (6)
1. the method for a quantum dot-labeled nano belt albumen, it is characterized in that, design a kind of fusion nano belt albumen of the random coil that contains Histidine, and add complementary polypeptide, make the random coil sequence of nano belt albumen be converted into the spiral curling of rigidity, make Histidine all in a spiral curling side, then spiral curling albumen is mixed with quantum dot, obtain quantum dot-labeled mixture.
2. the method for a kind of quantum dot-labeled nano belt albumen according to claim 1, it is characterized in that described fusion nano belt albumen is to add one section peptide sequence by albumen to be marked to form again, described peptide sequence comprises one section pep1, and the sequence of pep1 is HK VAQLKHE NQALEHE VASLEHK VSAL.
3. the method for a kind of quantum dot-labeled nano belt albumen according to claim 2, is characterized in that the complementary polypeptide added is pep2, and sequence is KEVQALEEK NAQLKEK VSALKKE VASLE.
4. the method for a kind of quantum dot-labeled nano belt albumen according to claim 3, is characterized in that described peptide sequence is formed by connecting by proline(Pro) by 5 pep1 sequences; The complementary polypeptide added is 5 pep2.
5. the method for a kind of quantum dot-labeled nano belt albumen according to claim 1, it is characterized in that: described quantum dot is the quantum dot that contains Zn.
6. the method for a kind of quantum dot-labeled nano belt albumen according to claim 1, it is characterized in that: the quantum dot that contains Zn is CdSe/ZnS, CdTe/ZnS, CdSe/ZnSe or CdTe/ZnSe.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108444876A (en) * | 2018-03-09 | 2018-08-24 | 国家纳米科学中心 | A kind of assay method of nano grain surface adhesion protein ligand state |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011050039A2 (en) * | 2009-10-22 | 2011-04-28 | The Government Of The United States Of America, As Represented By The Secretary Of The Navy | Modular functional peptides for the intracellular delivery of nanoparticles |
| CN102924564A (en) * | 2012-11-02 | 2013-02-13 | 常州大学 | Method for labeling protein through quantum dots |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011050039A2 (en) * | 2009-10-22 | 2011-04-28 | The Government Of The United States Of America, As Represented By The Secretary Of The Navy | Modular functional peptides for the intracellular delivery of nanoparticles |
| CN102924564A (en) * | 2012-11-02 | 2013-02-13 | 常州大学 | Method for labeling protein through quantum dots |
Non-Patent Citations (2)
| Title |
|---|
| KIM E. SAPSFORD等: "Kinetics of Metal-Affinity Driven Self-Assembly between Proteins or Peptides and CdSe-ZnS Quantum Dots", 《J. PHYS. CHEM. C》 * |
| 柳旭等: "水溶性ZnO量子点与生物分子相互作用的研究", 《稀有金属》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108444876A (en) * | 2018-03-09 | 2018-08-24 | 国家纳米科学中心 | A kind of assay method of nano grain surface adhesion protein ligand state |
| CN108444876B (en) * | 2018-03-09 | 2020-06-16 | 国家纳米科学中心 | Method for determining state of protein ligand adsorbed on surface of nanoparticle |
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