Citation: Sha Jingjie, Xu Bing, Chen Yunfei, Yang Yanjing. Experimental Research of Protein Translocation Using Solid-state Nanopore[J]. Acta Chimica Sinica, ;2017, 75(11): 1121-1125. doi: 10.6023/A17060271 shu

Experimental Research of Protein Translocation Using Solid-state Nanopore

Sha Jingjie ^a,b,, ,
Xu Bing ^a,b ,
Chen Yunfei ^a,b ,
Yang Yanjing ^c,d

a.
Jiangsu Key Laboratory for Design and Manufacture of Micro-Nano Biomedical Instruments, Southeast University, Nanjing 211189
b.
School of Mechanical Engineering, Southeast University, Nanjing 211189
c.
Jiangsu Key Laboratory of Oral Diseases, Nanjing Medical University, Nanjing 210029
d.
Department of Preventive Dentistry, Affiliated Hospital of Stomatology, Nanjing Medical University, Nanjing 210029

Corresponding author: Sha Jingjie, major212@seu.edu.cn
Received Date: 16 June 2017
Available Online: 7 November 2017
Fund Project: the National Natural Science Foundation of China 51675101the National Natural Science Foundation of China 51375092the Fundamental Research Funds for the Central Universities 2242015R30002the National Natural Science Foundation of China 51435003Project supported by the National Natural Science Foundation of China (Nos. 51375092, 51435003 and 51675101) and the Fundamental Research Funds for the Central Universities (No. 2242015R30002)

Figures(5)

For proteins' diverse range of structural and functional features, they are important populations of biomolecules within organisms. Common methods to detect proteins are with the help of fluorescence or enzyme. Due to the advantages like lable-free and single-molecule detection, nanopore technology provides a novel platform for proteins' characterization. In this experiment, the patch clamp amplifier is used to apply the voltage and acquire the tiny current blockage. The Si₃N₄ membrane drilled with a nanopore separated the buffer solution into two sides:cis and trans. When the voltage applied into the buffer solution, charged proteins would been driven through the pore from one side to the other. Then, a series of current blockages could be obtained. By analysing these data, the size and conformation of the biomolecules could be acquired. In this paper, we using solid-state nanopore detected single protein and protein-protein complexes. The nanopore was characterized firstly. Then, both the applied voltage and the pH of the electrolyte solution were regulated. Under the low voltage, the sample proteins could be regarded as a rigid spheroid, and the dwell time is decreased with the voltage increasing. It was found that, the charges carried by proteins could be improved by higher pH of buffer solution, so that the dwell time would been shortened. Furthermore, based on the high specific between the antigen and antibody which are proteins, the translocation events before and after the addition of specific antigen into the solution with antibody were compared. Results showed that the relative current drop of the complex is larger than the pure antibody, implying that the antigen has been bound into the antibody. Due to the difference of excluded volume, the antibody and antigen-antibody complexes could be distinguished by the solid-state nanopore. In the future, the nanopore technology is promising to be applied into the recognition of multiply proteins and protein-protein interaction.
- Bovine serum Albumin (BSA),
- antigen,
- antibody,
- solid-state nanopore,
- single-molecule detection
1. [1]
  Wang, J.-Y. Biochemistry 1, Higher Education Press, Beijing, 2002, pp. 123~124.
2. [2]
  Li, C.-Y. Basic Immunology, Science Press, Beijing, 2012, pp. 35~39 (in Chinese).
3. [3]
  Waduge, P.; Hu, R.; Bandarkar, P.; Yamazaki, H.; Cressiot, B.; Zhao, Q.; Whitford, P. C.; Wanunu, M. ACS Nano 2017, 11, 5706. doi: 10.1021/acsnano.7b01212
4. [4]
  Zhang, J.-Y.; Zhou, X.-Y.; Zhou, M.; Jia, H.-X. Acta Chim. Sinica 2016, 74, 513(in Chinese). doi: 10.3969/j.issn.0253-2409.2016.05.001
5. [5]
  Deamer, D.; Akeson, M.; Branton, D. Nat. Biotechnol. 2016, 34, 518. doi: 10.1038/nbt.3423
6. [6]
  Dekker, C. Nat. Nanotechnol. 2007, 2, 209. doi: 10.1038/nnano.2007.27
7. [7]
  Howorka, S.; Cheley, S.; Bayley, H. Nat. Biotechnol. 2001, 19, 636. doi: 10.1038/90236
8. [8]
  Meller, A.; Nivon, L.; Branton, D. Phys. Rev. Lett. 2001, 86, 3435. doi: 10.1103/PhysRevLett.86.3435
9. [9]
  Nakane, J. J.; Akeson, M.; Marziali, A. J. Phys.-Condens. Matter 2003, 15, 1365. doi: 10.1088/0953-8984/15/32/203
10. [10]
  Storm, A. J.; Storm, C.; Chen, J. H.; Zandbergen, H.; Joanny, J. F.; Dekker, C. Nano Lett. 2005, 5, 1193. doi: 10.1021/nl048030d
11. [11]
  Ying, Y. L.; Cao, C.; Long, Y. T. Analyst 2014, 139, 3826. doi: 10.1039/C4AN00706A
12. [12]
  Oukhaled, A.; Bacri, L.; Pastoriza-Gallego, M.; Betton, J. M.; Pelta, J. ACS Chem. Biol. 2012, 7, 1935. doi: 10.1021/cb300449t
13. [13]
  Kasianowicz, J. J.; Brandin, E.; Branton, D.; Deamer, D. W. Proceedings of the National Academy of Sciences of the United States of America 1996, 93, 13770. doi: 10.1073/pnas.93.24.13770
14. [14]
  Wendell, D.; Jing, P.; Geng, J.; Subramaniam, V.; Lee, T. J.; Montemagno, C.; Guo, P. Nat. Nanotechnol. 2009, 4, 765. doi: 10.1038/nnano.2009.259
15. [15]
  Cao, C.; Liao, D.-F.; Ying, Y.-L.; Long, Y.-T. Acta Chim. Sinica 2016, 74, 734(in Chinese).
16. [16]
  Ma, J.; Qiu, Y. H.; Yuan, Z. S.; Zhang, Y.; Sha, J. J.; Liu, L.; Sun, L. T.; Ni, Z. H.; Yi, H.; Li, D. Y.; Chen, Y. F. Phys. Rev. E 2015, 92, 022719. doi: 10.1103/PhysRevE.92.022719
17. [17]
  Wang, Y.; Yu, X.-F.; Liu, Y.-Y.; Xie, X.; Cheng, X.-L.; Huang, S.-M.; Wang, Z.-M. Acta Chim. Sinica 2014, 72, 378(in Chinese).
18. [18]
  Jiang, Y.-N.; Guo, W. Sci. Bull. 2015, 60, 491. doi: 10.1007/s11434-015-0739-6
19. [19]
  Guo, W.; Jiang, L. Sci. China Matter 2014, 57, 2. doi: 10.1007/s40843-014-0005-z
20. [20]
  Su, B.; Guo, W.; Jiang L. Small 2015, 11, 1072. doi: 10.1002/smll.v11.9-10
21. [21]
  Wu, L.-Z.; Liu, Y.-Q.; Liu, W.; Chen, D.; Chen, H. Acta Biophys. Sinica 2014, 5, 360(in Chinese).
22. [22]
  Farimani, A. B.; Heiranian, M.; Min, K.; Aluru, N. R. J. Phys. Chem. Lett. 2017, 8, 1670. doi: 10.1021/acs.jpclett.7b00385
23. [23]
  Freedman, K. J.; Jurgens, M.; Prabhu, A.; Ahn, C. W.; Jemth, P.; Edel, J. B.; Kim, M. J. Anal. Chem. 2011, 83, 5137. doi: 10.1021/ac2001725
24. [24]
  Niedzwiecki, D. J.; Grazul, J.; Movileanu, L. J. Am. Chem. Soc. 2010, 132, 10816. doi: 10.1021/ja1026858
25. [25]
  Ying, Y.-L.; Zhang, X.; Liu, Y.; Xue, M.-Z.; Li, H.-L.; Long, Y.-T. Acta Chim. Sinica 2013, 71, 44(in Chinese).
26. [26]
  Ying, Y.-L.; Long, Y.-T. Sci. Chi. Chem. 2017, 60, doi:10.1007/s11426-017-9082-5. doi: 10.1007/s11426-017-9082-5
27. [27]
  Reiner, J. E.; Balijepalli, A.; Robertson, J. W. F.; Campbell, J.; Suehle, J.; Kasianowicz, J. J. Chem. Rev. 2012, 112, 6431. doi: 10.1021/cr300381m
28. [28]
  Saleh, O. A.; Sohn, L. L. Proc. Natl. Acad. Sci. U. S. A. 2003, 100, 820. doi: 10.1073/pnas.0337563100
29. [29]
  Takakura, T.; Yanagi, I.; Goto, Y.; Ishige, Y.; Kohara, Y. Appl. Phys. Lett. 2016, 108, 123701. doi: 10.1063/1.4944641
30. [30]
  Alzghoul, S.; Hailat, M.; Zivanovic, S.; Que, L.; Shah, G. V. Biosens. Bioelectron. 2016, 77, 491. doi: 10.1016/j.bios.2015.10.006
31. [31]
  Kwak, D. K.; Chae, H.; Lee, M. K.; Ha, J. H.; Goyal, G.; Kim, M. J.; Kim, K. B.; Chi, S. W. Angew Chem., Int. Ed. 2016, 55, 5713. doi: 10.1002/anie.201511601
32. [32]
  Wang, S.; Haque, F.; Rychahou, P. G.; Evers, B. M.; Guo, P. ACS Nano 2013, 7, 9814. doi: 10.1021/nn404435v
33. [33]
  Han, A.; Creus, M.; Schurmann, G.; Linder, V.; Ward, T. R.; de Rooij, N. F.; Staufer, U. Anal. Chem. 2008, 80, 4651. doi: 10.1021/ac7025207
34. [34]
  Freedman, K. J.; Bastian, A. R.; Chaiken, I.; Kim, M. J. Small 2013, 9, 750. doi: 10.1002/smll.v9.5
35. [35]
  Peters, T. Adv. Protein. Chem. 1985, 37, 161. doi: 10.1016/S0065-3233(08)60065-0
36. [36]
  Li, J. L.; Talaga, D. S. J. Phys.-Condens. Matter 2010, 22, 454.
37. [37]
  Ling, D. Y.; Ling, X. S. J. Phys.-Condens. Matter 2013, 25, 375102. doi: 10.1088/0953-8984/25/37/375102
38. [38]
  Li, J. L.; Fologea, D.; Rollings, R.; Ledden, B. Protein and Peptide Letters. 2014, 21, 256. doi: 10.2174/09298665113209990077
39. [39]
  Deblois, R. W.; Bean, C. P. Rev. Sci. Instrum. 1970, 41, 909. doi: 10.1063/1.1684724
40. [40]
  Han, A. P.; Schurmann, G.; Mondin, G.; Bitterli, R. A.; Hegelbach, N. G.; de Rooij, N. F.; Staufer, U. Appl. Phys. Lett. 2006, 88, 093901. doi: 10.1063/1.2180868
41. [41]
  Larkin, J.; Henley, R. Y.; Muthukumar, M.; Rosenstein, J. K.; Wanunu, M. Biophys. J. 2014, 106, 696. doi: 10.1016/j.bpj.2013.12.025
1. [1]
  Zeyu XU , Anlei DANG , Bihua DENG , Xiaoxin ZUO , Yu LU , Ping YANG , Wenzhu YIN . Evaluation of the efficacy of graphene oxide quantum dots as an ovalbumin delivery platform and adjuvant for immune enhancement. Chinese Journal of Inorganic Chemistry, 2024, 40(6): 1065-1078. doi: 10.11862/CJIC.20240099
2. [2]
  Jingwen Wang , Minghao Wu , Xing Zuo , Yaofeng Yuan , Yahao Wang , Xiaoshun Zhou , Jianfeng Yan . Advances in the Application of Electrochemical Regulation in Investigating the Electron Transport Properties of Single-Molecule Junctions. University Chemistry, 2025, 40(3): 291-301. doi: 10.12461/PKU.DXHX202406023
3. [3]
  Chunmei GUO , Weihan YIN , Jingyi SHI , Jianhang ZHAO , Ying CHEN , Quli FAN . Facile construction and peroxidase-like activity of single-atom platinum nanozyme. Chinese Journal of Inorganic Chemistry, 2024, 40(9): 1633-1639. doi: 10.11862/CJIC.20240162
4. [4]
  Yang YANG , Pengcheng LI , Zhan SHU , Nengrong TU , Zonghua WANG . Plasmon-enhanced upconversion luminescence and application of molecular detection. Chinese Journal of Inorganic Chemistry, 2024, 40(5): 877-884. doi: 10.11862/CJIC.20230440
5. [5]
  Yuanpei ZHANG , Jiahong WANG , Jinming HUANG , Zhi HU . Preparation of magnetic mesoporous carbon loaded nano zero-valent iron for removal of Cr(Ⅲ) organic complexes from high-salt wastewater. Chinese Journal of Inorganic Chemistry, 2024, 40(9): 1731-1742. doi: 10.11862/CJIC.20240077
6. [6]
  Hong LI , Xiaoying DING , Cihang LIU , Jinghan ZHANG , Yanying RAO . Detection of iron and copper ions based on gold nanorod etching colorimetry. Chinese Journal of Inorganic Chemistry, 2024, 40(5): 953-962. doi: 10.11862/CJIC.20230370
7. [7]
  Jinghan ZHANG , Guanying CHEN . Progress in the application of rare-earth-doped upconversion nanoprobes in biological detection. Chinese Journal of Inorganic Chemistry, 2024, 40(12): 2335-2355. doi: 10.11862/CJIC.20240249
8. [8]
  Yongjie ZHANG , Bintong HUANG , Yueming ZHAI . Research progress of formation mechanism and characterization techniques of protein corona on the surface of nanoparticles. Chinese Journal of Inorganic Chemistry, 2024, 40(12): 2318-2334. doi: 10.11862/CJIC.20240247
9. [9]
  Yuhao SUN , Qingzhe DONG , Lei ZHAO , Xiaodan JIANG , Hailing GUO , Xianglong MENG , Yongmei GUO . Synthesis and antibacterial properties of silver-loaded sod-based zeolite. Chinese Journal of Inorganic Chemistry, 2024, 40(4): 761-770. doi: 10.11862/CJIC.20230169
10. [10]
  Junjie Zhang , Yue Wang , Qiuhan Wu , Ruquan Shen , Han Liu , Xinhua Duan . Preparation and Selective Separation of Lightweight Magnetic Molecularly Imprinted Polymers for Trace Tetracycline Detection in Milk. University Chemistry, 2024, 39(5): 251-257. doi: 10.3866/PKU.DXHX202311084
11. [11]
  Liwei Wang , Guangran Ma , Li Wang , Fugang Xu . A Comprehensive Analytical Chemistry Experiment: Colorimetric Detection of Vitamin C Using Nanozyme and Smartphone. University Chemistry, 2024, 39(8): 255-262. doi: 10.3866/PKU.DXHX202312094
12. [12]
  Yufang GAO , Nan HOU , Yaning LIANG , Ning LI , Yanting ZHANG , Zelong LI , Xiaofeng LI . Nano-thin layer MCM-22 zeolite: Synthesis and catalytic properties of trimethylbenzene isomerization reaction. Chinese Journal of Inorganic Chemistry, 2024, 40(6): 1079-1087. doi: 10.11862/CJIC.20240036
13. [13]
  Pingwei Wu . Application of Diamond Software in Simplex Teaching. University Chemistry, 2024, 39(3): 118-121. doi: 10.3866/PKU.DXHX202311043
14. [14]
  Tao Jiang , Yuting Wang , Lüjin Gao , Yi Zou , Bowen Zhu , Li Chen , Xianzeng Li . Experimental Design for the Preparation of Composite Solid Electrolytes for Application in All-Solid-State Batteries: Exploration of Comprehensive Chemistry Laboratory Teaching. University Chemistry, 2024, 39(2): 371-378. doi: 10.3866/PKU.DXHX202308057
15. [15]
  Wenqi Gao , Xiaoyan Fan , Feixiang Wang , Zhuojun Fu , Jing Zhang , Enlai Hu , Peijun Gong . Exploring Nernst Equation Factors and Applications of Solid Zinc-Air Battery. University Chemistry, 2024, 39(5): 98-107. doi: 10.3866/PKU.DXHX202310026
16. [16]
  Rui Li , Huan Liu , Yinan Jiao , Shengjian Qin , Jie Meng , Jiayu Song , Rongrong Yan , Hang Su , Hengbin Chen , Zixuan Shang , Jinjin Zhao . 卤化物钙钛矿的单双向离子迁移. Acta Physico-Chimica Sinica, 2024, 40(11): 2311011-. doi: 10.3866/PKU.WHXB202311011
17. [17]
  Di Yang , Jiayi Wei , Hong Zhai , Xin Wang , Taiming Sun , Haole Song , Haiyan Wang . Rapid Detection of SARS-CoV-2 Using an Innovative “Magic Strip”. University Chemistry, 2024, 39(4): 373-381. doi: 10.3866/PKU.DXHX202312023
18. [18]
  Mingyang Men , Jinghua Wu , Gaozhan Liu , Jing Zhang , Nini Zhang , Xiayin Yao . 液相法制备硫化物固体电解质及其在全固态锂电池中的应用. Acta Physico-Chimica Sinica, 2025, 41(1): 2309019-. doi: 10.3866/PKU.WHXB202309019
19. [19]
  Fei Xie , Chengcheng Yuan , Haiyan Tan , Alireza Z. Moshfegh , Bicheng Zhu , Jiaguo Yu . d带中心调控过渡金属单原子负载COF吸附O₂的理论计算研究. Acta Physico-Chimica Sinica, 2024, 40(11): 2407013-. doi: 10.3866/PKU.WHXB202407013
20. [20]
  Yang WANG , Xiaoqin ZHENG , Yang LIU , Kai ZHANG , Jiahui KOU , Linbing SUN . Mn single-atom catalysts based on confined space: Fabrication and the electrocatalytic oxygen evolution reaction performance. Chinese Journal of Inorganic Chemistry, 2024, 40(11): 2175-2185. doi: 10.11862/CJIC.20240165

Get Citation

PDF

XML

Metrics

PDF Downloads(11)
Abstract views(2302)
HTML views(240)

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142