Expression, Purification and Characterization of Recombinant Disintegrin from Gloydius Brevicaudus Venom in Escherichia Coli

IF 1.9 4区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY The Protein Journal Pub Date : 2024-05-11 DOI:10.1007/s10930-024-10198-w

Yinxiang Lan, Xiuliang Qiu, Yunlu Xu

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Abstract

Disintegrins, a family of snake venom protein, which are capable of modulating the activity of integrins that play a fundamental role in the regulation of many physiological and pathological processes. The main purpose of this study is to obtain the recombinant disintegrin (r-DI) and evaluate its biological activity. In this study, we explored a high-level expression prokaryotic system and purification strategy for r-DI. Then, r-DI was treated to assay effects on cell growth, migration, and invasion. The affinity for the interactions of r-DI with integrin was determined using Surface plasmon resonance (SPR) analyses. The r-DI can be expressed in Escherichia coli and purified by one-step chromatography. The r-DI can inhibit B16F10 cells proliferation, migration, and invasion. Also, we found that r-DI could interact with the integrin αIIbβ3 (GPIIb/IIIa). The r-DI can be expressed, purified, characterized through functional assays, and can also maintain strong biological activities. Thus, this study showed potential therapeutic effects of r-DI for further functional and structural studies.

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在大肠杆菌中表达、纯化和表征来自 Gloydius Brevicaudus 毒液的重组崩解素。

崩解素是蛇毒蛋白的一个家族，能够调节整合素的活性，而整合素在许多生理和病理过程的调节中发挥着重要作用。本研究的主要目的是获得重组崩解素（r-DI）并评估其生物活性。在这项研究中，我们探索了一种高水平的原核表达系统和 r-DI 的纯化策略。然后，对 r-DI 进行处理，检测其对细胞生长、迁移和侵袭的影响。利用表面等离子体共振（SPR）分析确定了 r-DI 与整合素相互作用的亲和力。r-DI 可以在大肠杆菌中表达，并通过一步色谱法纯化。r-DI 可抑制 B16F10 细胞的增殖、迁移和侵袭。此外，我们还发现 r-DI 可与整合素 αIIbβ3 （GPIIb/IIIa）相互作用。r-DI 可以表达、纯化，并通过功能测试进行表征，还能保持很强的生物活性。因此，这项研究显示了 r-DI 的潜在治疗作用，可用于进一步的功能和结构研究。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

The Protein Journal 生物-生化与分子生物学

CiteScore

5.20

自引率

0.00%

发文量

审稿时长

12 months

期刊介绍： The Protein Journal (formerly the Journal of Protein Chemistry) publishes original research work on all aspects of proteins and peptides. These include studies concerned with covalent or three-dimensional structure determination (X-ray, NMR, cryoEM, EPR/ESR, optical methods, etc.), computational aspects of protein structure and function, protein folding and misfolding, assembly, genetics, evolution, proteomics, molecular biology, protein engineering, protein nanotechnology, protein purification and analysis and peptide synthesis, as well as the elucidation and interpretation of the molecular bases of biological activities of proteins and peptides. We accept original research papers, reviews, mini-reviews, hypotheses, opinion papers, and letters to the editor.