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

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

doi:10.1007/s10930-024-10244-7

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

Protein J. 2024 Dec 19. doi: 10.1007/s10930-024-10244-7. Online ahead of print.

Authors

Rayees Ahmad Lone^#^{1

2}, Bhupendra Kumar^#^{1

2}, Mohd Kashif³, Shafquat Fakhrah¹, Tofan Kumar Rout⁴, Sahabjada Siddiqui⁵, Rojalin Pattanayak⁶, Pradhyumna Kumar Singh^{1

2}, Chandra Sekhar Mohanty^{7

8}

Affiliations

¹ Plant Genetic Resources and Improvement Division, CSIR-National Botanical Research Institute, Lucknow, Uttar Pradesh, India.
² Academy of Scientific and Innovative Research, Ghaziabad, Uttar Pradesh, India.
³ Babasaheb Bhimrao Ambedkar University, Lucknow, Uttar Pradesh, India.
⁴ Sophisticated Analytical Instrument Facility and Research, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh, India.
⁵ Department of Biotechnology, Era University, Lucknow, Uttar Pradesh, India.
⁶ College of Basic Science and Humanities, Odisha University of Agriculture and Technology, Bhubaneswar, Odisha, India.
⁷ Plant Genetic Resources and Improvement Division, CSIR-National Botanical Research Institute, Lucknow, Uttar Pradesh, India. cs.mohanti@nbri.res.in.
⁸ Academy of Scientific and Innovative Research, Ghaziabad, Uttar Pradesh, India. cs.mohanti@nbri.res.in.

^# Contributed equally.

PMID: 39702856
DOI: 10.1007/s10930-024-10244-7

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.

Keywords: Anticancer; Kunitz-type; Root-tuber; Trypsin inhibitor; Winged bean.