基于表面等离子体共振(SPR)技术实时分析RGD基序与整联蛋白的相互作用

doi:10.11843/j.issn.0366-6964.2021.027

Abstract

Abstract: This experiment aims to study the binding specificity of RGD (Arg-Gly-Asp) motif on the structural protein VP1 of foot-and-mouth disease virus (FMDV) to host cell surface receptor integrin. In this paper, the Biacore 3000 system based on surface plasmon resonance (SPR) was used to study the binding specificity of RGD motif with the α_vβ₆ extracellular domain, α_v extracellular domain and β₆ extracellular domain, respectively. Firstly, the domains of integrin bound to RGD Motif were screened by binding experiments, and then the kinetic analysis of integrin bound to RGD Motif was carried out. The results showed that the synthesized RGD motif was bound to the α_vβ₆ extracellular domain, and the kinetic constants K_a, K_d, and K_D were 42.3 M^-1s^-1, 3.1×10^-4s^-1, and 7.33×10^-6M respectively. It also binds to the α_v extracellular domain, the kinetic constants were 21.8 M^-1s^-1, 2.13×10^-4s^-1,and 9.79×10^-5M respectively. There was almost no binding to the extracellular domain of the β₆ subunit. The results showed that the binding of RGD to the extracellular domain of α_vβ₆ was faster and stronger than that of the extracellular domain of α_v subunit. This study will provide a reference for further study of the interaction between FMDV and host cell surface receptors.

Key words: Biacore 3000, FMDV, RGD motif, surface plasmon resonance, integrin, kinetics

CLC Number:

S855.3

LI Pengfei, GONG Zhenli, DU Xiaohua, JIANG Tao. Real-time Analysis of the Interaction between RGD Motif and Integrin Based on Surface Plasmon Resonance (SPR)[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(1): 256-261.

References 28

[1]	PATON D J, GUBBINS S, KING D P. Understanding the transmission of foot-and-mouth disease virus at different scales[J]. Curr Opin Virol, 2018, 28:85-91.
[2]	ABUBAKAR M, MANZOOR S, AHMED A. Interplay of foot and mouth disease virus with cell-mediated and humoral immunity of host[J]. Rev Med Virol, 2018, 28(2):e1966.
[3]	JAMAL S M, BELSHAM G J. Molecular epidemiology, evolution and phylogeny of foot-and-mouth disease virus[J]. Infect Genet Evol, 2018, 59:84-98.
[4]	VASQUEZ C, DENOYA C D, LA TORRE J, et al. Structure of foot-and-mouth disease virus capsid[J]. Virology, 1979, 97(1):195-200.
[5]	SOBRINO F, SÁIZ M, JIMÉNEZ-CLAVERO M A, et al. Foot-and-mouth disease virus:a long known virus, but a current threat[J]. Vet Res, 2001, 32(1):1-30.
[6]	BERRYMAN S, CLARK S, MONAGHAN P, et al. Early events in integrin αvβ6-mediated cell entry of foot-and-mouth disease virus[J]. J Virol, 2005, 79(13):8519-8534.
[7]	LIAN K Q, YANG F, ZHU Z X, et al. The VP1 S154D mutation of type Asia 1 foot-and-mouth disease virus enhances viral replication and pathogenicity[J]. Infect Genet Evol, 2016, 39:113-119.
[8]	HYNES R O. Integrins:a family of cell surface receptors[J]. Cell, 1987, 48(4):549-554.
[9]	GAHMBERG C G, GRÖNHOLM M, MADHAVAN S, et al. Regulation of cell adhesion:a collaborative effort of integrins, their ligands, cytoplasmic actors, and phosphorylation[J]. Q Rev Biophys, 2019, 52:e10.
[10]	KOIVISTO L, BI J R, HÄKKINEN L, et al. Integrin αvβ6:Structure, function and role in health and disease[J]. Int J Biochem Cell Biol, 2018, 99:186-196.
[11]	HUSSEIN H A M, WALKER L R, ABDEL-RAOUF U M, et al. Beyond RGD:virus interactions with integrins[J]. Arch Virol, 2015, 160(11):2669-2681.
[12]	WANG G X, WANG Y H, SHANG Y J, et al. How foot-and-mouth disease virus receptor mediates foot-and-mouth disease virus infection[J]. Virol J, 2015, 12:9.
[13]	SHIMMON G, WOOD B A, MORRIS A, et al. Truncated bovine integrin alpha-v/beta-6 as a universal capture ligand for FMD diagnosis[J]. PLoS One, 2016, 11(8):e0160696.
[14]	BISWAL J K, MOHAPATRA J K, BISHT P, et al. A positively charged lysine residue at VP2131 position allows for the enhanced adaptability of foot-and-mouth disease virus serotype A in BHK-21 cells[J]. Biologicals, 2015, 43(1):71-78.
[15]	CHAMBERLAIN K, FOWLER V L, BARNETT P V, et al. Identification of a novel cell culture adaptation site on the capsid of foot-and-mouth disease virus[J]. J Gen Virol, 2015, 96(9):2684-2692.
[16]	LUO B H, CARMAN C V, SPRINGER T A. Structural basis of integrin regulation and signaling[J]. Annu Rev Immunol, 2007, 25:619-647.
[17]	安娜, 郑子繁, 董美, 等. 基于表面等离子体共振的核酸传感器研究进展[J]. 中国农业科技导报, 2019, 21(5):55-61.AN N, ZHENG Z F, DONG M, et al. Research progress on nucleic acid sensors based on surface plasmon resonance[J]. Journal of Agricultural Science and Technology, 2019, 21(5):55-61. (in Chinese)
[18]	BERRYMAN J. Frequently asked questions patient simulator mannequin[J]. Colo Nurse, 2005, 105(3):28.
[19]	STEWART P L, NEMEROW G R. Cell integrins:commonly used receptors for diverse viral pathogens[J]. Trends Microbiol, 2007, 15(11):500-507.
[20]	BAI X W, BAO H F, LI P H, et al. Effects of two amino acid substitutions in the capsid proteins on the interaction of two cell-adapted PanAsia-1 strains of foot-and-mouth disease virus serotype O with heparan sulfate receptor[J]. Virol J, 2014, 11:132.
[21]	BAI X W, BAO H F, LI P H, et al. Engineering responses to amino acid substitutions in the VP0-and VP3-coding regions of panasia-1 strains of foot-and-mouth disease virus serotype O[J]. J Virol, 2019, 93(7):e02278-18.
[22]	BISWAL J K, DAS B, SHARMA G K, et al. Role of a single amino acid substitution of VP3 H142D for increased acid resistance of foot-and-mouth disease virus serotype A[J]. Virus Genes, 2016, 52(2):235-243.
[23]	SINGH I, DEB R, KUMAR S, et al. Deciphering foot-and-mouth disease (FMD) virus-host tropism[J]. J Biomol Struct Dyn, 2019, 37(18):4779-4789.
[24]	DICARA D, BURMAN A, CLARK S, et al. Foot-and-mouth disease virus forms a highly stable, EDTA-resistant complex with its principal receptor, integrin alphavbeta6:implications for infectiousness[J]. J Virol, 2008, 82(3):1537-1546.
[25]	VAN AGTHOVEN J F, XIONG J P, ALONSO J L, et al. Structural basis for pure antagonism of integrin αVβ3 by a high-affinity form of fibronectin[J]. Nat Struct Mol Biol, 2014, 21(4):383-388.
[26]	DONG X C, HUDSON N E, LU C F, et al. Structural determinants of integrin β-subunit specificity for latent TGF-β[J]. Nat Struct Mol Biol, 2014, 21(12):1091-1096.
[27]	TIWARI S, ASKARI J A, HUMPHRIES M J, et al. Divalent cations regulate the folding and activation status of integrins during their intracellular trafficking[J]. J Cell Sci, 2011, 124(10):1672-1680.
[28]	HALL E R, SLACK R J. The effect of divalent metal cations on the αv integrin binding site is ligand and integrin specific[J]. Biomed Pharmacother, 2019, 110:362-370.

Real-time Analysis of the Interaction between RGD Motif and Integrin Based on Surface Plasmon Resonance (SPR)

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