The Root-Tuber Trypsin Inhibitor of Winged Bean and Its Anti-cancerous Activity Against Osteosarcoma Cell-Line

IF 1.9 4区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

The Protein Journal Pub Date : 2024-12-19 DOI:10.1007/s10930-024-10244-7

Rayees Ahmad Lone, Bhupendra Kumar, Mohd. Kashif, Shafquat Fakhrah, Tofan Kumar Rout, Sahabjada Siddiqui, Rojalin Pattanayak, Pradhyumna Kumar Singh, Chandra Sekhar Mohanty

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Abstract

Trypsin inhibitor from the root-tuber of underutilized legume Winged bean (Psophocarpus tetragonolobus (L.) DC.) (WbT-TI) was purified using ion exchange chromatography followed by size-exclusion chromatography. The purified WbT-TI showed a molecular mass of 20,609 Da and an isoelectric point of 5.10. Ultraviolet circular dichroism (UV-CD) and intrinsic fluorescence reported, that WbT-TI interacts with trypsin. Domain-wise analysis of WbT-TI revealed it to belong to the Kunitz-type soybean trypsin inhibitor (STI) family with a specific β-trefoil fold. The sequence of WbT-TI showed 44% sequence coverage to acidic trypsin inhibitor from the seed of the same plant. Protein interaction similarity analysis (PIPSA) evaluated the electrostatic properties of WbT-TI and provided information about the interacting partners of trypsin inhibitors. The purified protein was quantified and tested for in vitro anticancer activity using 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay against the human osteosarcoma (MG-63) cell line. At 5 µg/ml of WbT-TI, the highest inhibition was seen. These studies may lead to the development of winged bean protease inhibitor-based preventive and therapeutic strategies for different kinds of cancers.

Graphical Abstract

Abstract Image

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豇豆块根胰蛋白酶抑制剂及其对骨肉瘤细胞的抗癌作用。

未充分利用的豆科植物飞豆根茎中胰蛋白酶抑制剂的研究DC.) （WbT-TI）经离子交换层析-排粒径层析纯化。纯化后的WbT-TI分子量为20609 Da，等电点为5.10。紫外圆二色性（UV-CD）和本征荧光研究表明WbT-TI与胰蛋白酶相互作用。WbT-TI的结构域分析显示它属于kunitz型大豆胰蛋白酶抑制剂（STI）家族，具有特异性的β-三叶折叠。WbT-TI对同一植物种子酸性胰蛋白酶抑制剂的序列覆盖率为44%。蛋白相互作用相似性分析（PIPSA）评价了WbT-TI的静电特性，并提供了胰蛋白酶抑制剂相互作用伙伴的信息。用3-(4,5-二甲基噻唑-2)-2,5-二苯基溴化四唑（MTT）法测定纯化蛋白对人骨肉瘤（MG-63）细胞系的体外抗癌活性。WbT-TI浓度为5µg/ml时，抑制作用最强。这些研究可能会导致以翼豆蛋白酶抑制剂为基础的不同类型癌症的预防和治疗策略的发展。

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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.