Multimeric Association of Purified Novel Bowman-Birk Inhibitor From the Medicinal Forage Legume Mucuna pruriens (L.) DC.

A Bowman-Birk protease, i.e., Mucuna pruriens trypsin inhibitor (MPTI), was purified from the seeds by 55.702-fold and revealed a single trypsin inhibitor on a zymogram with a specific activity of 202.31 TIU/mg of protein. On sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) under...

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Autores principales:	Jafar K. Lone, Mandapanda A. Lekha, Rajiv P. Bharadwaj, Fasil Ali, M. Arumugam Pillai, Shabir H. Wani, Jeshima Khan Yasin, K. S. Chandrashekharaiah
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	anti-inflammatory activity multimeric association Bowman-Birk inhibitor seed proteins Mucuna pruriens Plant culture SB1-1110
Acceso en línea:	https://doaj.org/article/694173b8d47d46a3a54f897c74248854
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spelling	oai:doaj.org-article:694173b8d47d46a3a54f897c742488542021-12-01T00:52:21ZMultimeric Association of Purified Novel Bowman-Birk Inhibitor From the Medicinal Forage Legume Mucuna pruriens (L.) DC.1664-462X10.3389/fpls.2021.772046https://doaj.org/article/694173b8d47d46a3a54f897c742488542021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fpls.2021.772046/fullhttps://doaj.org/toc/1664-462XA Bowman-Birk protease, i.e., Mucuna pruriens trypsin inhibitor (MPTI), was purified from the seeds by 55.702-fold and revealed a single trypsin inhibitor on a zymogram with a specific activity of 202.31 TIU/mg of protein. On sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) under non-reducing conditions, the protease trypsin inhibitor fraction [i.e., trypsin inhibitor non-reducing (TINR)] exhibited molecular weights of 74 and 37 kDa, and under reducing conditions [i.e., trypsin inhibitor reducing (TIR)], 37 and 18 kDa. TINR-37 revealed protease inhibitor activity on native PAGE and 37 and 18 kDa protein bands on SDS–PAGE. TINR-74 showed peaks corresponding to 18.695, 37.39, 56.085, and 74.78 kDa on ultra-performance liquid chromatography (UPLC) coupled with electrospray ionization/quadrupole time-of-flight-mass spectrometry (ESI/QTOF-MS). Similarly, TINR-37 displayed 18.695 and 37.39 kDa peaks. Furthermore, TIR-37 and TIR-18 exhibited peaks corresponding to 37.39 and 18.695 kDa. Multiple peaks observed by the UPLC-ESI/QTOF analysis revealed the multimeric association, confirming the characteristic and functional features of Bowman-Birk inhibitors (BBIs). The multimeric association helps to achieve more stability, thus enhancing their functional efficiency. MPTI was found to be a competitive inhibitor which again suggested that it belongs to the BBI family of inhibitors, displayed an inhibitor constant of 1.3 × 10–6 M, and further demonstrates potent anti-inflammatory activity. The study provided a comprehensive basis for the identification of multimeric associates and their therapeutic potential, which could elaborate the stability and functional efficiency of the MPTI in the native state from M. pruriens.Jafar K. LoneMandapanda A. LekhaRajiv P. BharadwajFasil AliM. Arumugam PillaiShabir H. WaniJeshima Khan YasinK. S. ChandrashekharaiahFrontiers Media S.A.articleanti-inflammatory activitymultimeric associationBowman-Birk inhibitorseed proteinsMucuna pruriensPlant cultureSB1-1110ENFrontiers in Plant Science, Vol 12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	anti-inflammatory activity multimeric association Bowman-Birk inhibitor seed proteins Mucuna pruriens Plant culture SB1-1110
spellingShingle	anti-inflammatory activity multimeric association Bowman-Birk inhibitor seed proteins Mucuna pruriens Plant culture SB1-1110 Jafar K. Lone Mandapanda A. Lekha Rajiv P. Bharadwaj Fasil Ali M. Arumugam Pillai Shabir H. Wani Jeshima Khan Yasin K. S. Chandrashekharaiah Multimeric Association of Purified Novel Bowman-Birk Inhibitor From the Medicinal Forage Legume Mucuna pruriens (L.) DC.
description	A Bowman-Birk protease, i.e., Mucuna pruriens trypsin inhibitor (MPTI), was purified from the seeds by 55.702-fold and revealed a single trypsin inhibitor on a zymogram with a specific activity of 202.31 TIU/mg of protein. On sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) under non-reducing conditions, the protease trypsin inhibitor fraction [i.e., trypsin inhibitor non-reducing (TINR)] exhibited molecular weights of 74 and 37 kDa, and under reducing conditions [i.e., trypsin inhibitor reducing (TIR)], 37 and 18 kDa. TINR-37 revealed protease inhibitor activity on native PAGE and 37 and 18 kDa protein bands on SDS–PAGE. TINR-74 showed peaks corresponding to 18.695, 37.39, 56.085, and 74.78 kDa on ultra-performance liquid chromatography (UPLC) coupled with electrospray ionization/quadrupole time-of-flight-mass spectrometry (ESI/QTOF-MS). Similarly, TINR-37 displayed 18.695 and 37.39 kDa peaks. Furthermore, TIR-37 and TIR-18 exhibited peaks corresponding to 37.39 and 18.695 kDa. Multiple peaks observed by the UPLC-ESI/QTOF analysis revealed the multimeric association, confirming the characteristic and functional features of Bowman-Birk inhibitors (BBIs). The multimeric association helps to achieve more stability, thus enhancing their functional efficiency. MPTI was found to be a competitive inhibitor which again suggested that it belongs to the BBI family of inhibitors, displayed an inhibitor constant of 1.3 × 10–6 M, and further demonstrates potent anti-inflammatory activity. The study provided a comprehensive basis for the identification of multimeric associates and their therapeutic potential, which could elaborate the stability and functional efficiency of the MPTI in the native state from M. pruriens.
format	article
author	Jafar K. Lone Mandapanda A. Lekha Rajiv P. Bharadwaj Fasil Ali M. Arumugam Pillai Shabir H. Wani Jeshima Khan Yasin K. S. Chandrashekharaiah
author_facet	Jafar K. Lone Mandapanda A. Lekha Rajiv P. Bharadwaj Fasil Ali M. Arumugam Pillai Shabir H. Wani Jeshima Khan Yasin K. S. Chandrashekharaiah
author_sort	Jafar K. Lone
title	Multimeric Association of Purified Novel Bowman-Birk Inhibitor From the Medicinal Forage Legume Mucuna pruriens (L.) DC.
title_short	Multimeric Association of Purified Novel Bowman-Birk Inhibitor From the Medicinal Forage Legume Mucuna pruriens (L.) DC.
title_full	Multimeric Association of Purified Novel Bowman-Birk Inhibitor From the Medicinal Forage Legume Mucuna pruriens (L.) DC.
title_fullStr	Multimeric Association of Purified Novel Bowman-Birk Inhibitor From the Medicinal Forage Legume Mucuna pruriens (L.) DC.
title_full_unstemmed	Multimeric Association of Purified Novel Bowman-Birk Inhibitor From the Medicinal Forage Legume Mucuna pruriens (L.) DC.
title_sort	multimeric association of purified novel bowman-birk inhibitor from the medicinal forage legume mucuna pruriens (l.) dc.
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/694173b8d47d46a3a54f897c74248854
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Multimeric Association of Purified Novel Bowman-Birk Inhibitor From the Medicinal Forage Legume Mucuna pruriens (L.) DC.

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