Expression of a heptelidic acid-insensitive recombinant GAPDH from Trichoderma virens, and its biochemical and biophysical characterization.


Journal

Protein expression and purification
ISSN: 1096-0279
Titre abrégé: Protein Expr Purif
Pays: United States
ID NLM: 9101496

Informations de publication

Date de publication:
11 2020
Historique:
received: 07 04 2020
revised: 08 06 2020
accepted: 26 06 2020
pubmed: 19 7 2020
medline: 29 1 2021
entrez: 19 7 2020
Statut: ppublish

Résumé

Trichoderma virens genome harbors two isoforms of GAPDH, one (gGPD) involved in glycolysis and the other one (vGPD) in secondary metabolism. vGPD is expressed as part of the "vir" cluster responsible for the biosynthesis of volatile sesquiterpenes. The secondary metabolism-associated GAPDH is tolerant to the anti-cancer metabolite heptelidic acid (HA), produced by T. virens. Characterizing the HA-tolerant form of GAPDH, thus has implications in cancer therapy. In order to get insight into the mechanism of HA-tolerance of vGPD, we have purified recombinant form of this protein. The protein displays biochemical and biophysical characteristics analogous to the gGPD isoform. It exists as a tetramer with Tm of about 56.5 °C, and displays phosphorylation enzyme activity with Km and Kcat of 0.38 mM and 2.55 sec

Identifiants

pubmed: 32681951
pii: S1046-5928(20)30288-6
doi: 10.1016/j.pep.2020.105697
pii:
doi:

Substances chimiques

Fungal Proteins 0
Recombinant Proteins 0
Sesquiterpenes 0
heptelidic acid 57710-57-3
Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) EC 1.2.1.12

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

105697

Informations de copyright

Copyright © 2020 Elsevier Inc. All rights reserved.

Auteurs

Shikha Pachauri (S)

Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Trombay, Mumbai, 400085, India; Homi Bhabha National Institute, Anushaktinagar, Mumbai, 400094, India.

Gagan D Gupta (GD)

Homi Bhabha National Institute, Anushaktinagar, Mumbai, 400094, India; Radiation Biology & Health Sciences Division, Bhabha Atomic Research Centre, Trombay, Mumbai, 400085, India.

Prasun K Mukherjee (PK)

Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Trombay, Mumbai, 400085, India; Homi Bhabha National Institute, Anushaktinagar, Mumbai, 400094, India. Electronic address: prasunmukherjee1@gmail.com.

Vinay Kumar (V)

Homi Bhabha National Institute, Anushaktinagar, Mumbai, 400094, India. Electronic address: vkbhatia@gmail.com.

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Classifications MeSH