title: Purification and characterization of glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) from pea seeds.
abstract: Glyceraldehyde-3-phosphate dehydrogenase [GAPDH, NAD + oxidoreductase (phosphorylating) 1.2.1.12] catalyzes the conversion of glyceraldehyde-3-phosphate to 1,3-bisphosphoglycerate coupled with the reduction of NAD(+) to NADH. In addition to its role in glycolysis, this enzyme has numerous alternate functions, in both prokaryotes and eukaryotes. In plants, additional functions have been reported from multiple species including Pisum sativum. A recent study has identified that GAPDH may play an important role in seed ageing and programmed cell death. Despite this the existing purification protocols are almost 40 years old, and only partial characterization of the enzyme has been reported. In the current study, we report a modified method for purification of enzymatically active pea seed GAPDH along with the characterization of the enzyme. Using 2D gel electrophoresis our study also demonstrates that pea seeds contain four isoforms of NAD(+) dependent GAPDH.
authors.affiliation: National Institute of Pharmaceutical Education and Research, Phase X, Sector 67, SAS Nagar, Punjab, 160067, India.
authors.forename: Zahid
authors.initials: Z
authors.lastname: Gani
authors.affiliation: National Institute of Pharmaceutical Education and Research, Phase X, Sector 67, SAS Nagar, Punjab, 160067, India.
authors.forename: Vishant Mahendra
authors.initials: VM
authors.lastname: Boradia
authors.affiliation: National Institute of Pharmaceutical Education and Research, Phase X, Sector 67, SAS Nagar, Punjab, 160067, India.
authors.forename: Janaki
authors.initials: J
authors.lastname: Raghu Ram
authors.affiliation: National Institute of Pharmaceutical Education and Research, Phase X, Sector 67, SAS Nagar, Punjab, 160067, India.
authors.forename: Prashant Mohan
authors.initials: PM
authors.lastname: Suryavanshi
authors.affiliation: National Institute of Pharmaceutical Education and Research, Phase X, Sector 67, SAS Nagar, Punjab, 160067, India.
authors.forename: Pravinkumar
authors.initials: P
authors.lastname: Patil
authors.affiliation: Institute of Microbial Technology, Sector 39 A, Chandigarh, 160037, India.
authors.forename: Santosh
authors.initials: S
authors.lastname: Kumar
authors.affiliation: National Institute of Pharmaceutical Education and Research, Phase X, Sector 67, SAS Nagar, Punjab, 160067, India.
authors.forename: Ranvir
authors.initials: R
authors.lastname: Singh
authors.affiliation: Institute of Microbial Technology, Sector 39 A, Chandigarh, 160037, India.
authors.forename: Manoj
authors.initials: M
authors.lastname: Raje
authors.affiliation: National Institute of Pharmaceutical Education and Research, Phase X, Sector 67, SAS Nagar, Punjab, 160067, India. Electronic address: chaaya@niper.ac.in.
authors.forename: Chaaya Iyengar
authors.initials: CI
authors.lastname: Raje
mesh_terms: D000818:Animals; D002645:Chickens; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry* / Q000302:isolation & purification*; D018532:Pisum sativum / Q000201:enzymology*; D010940:Plant Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D011817:Rabbits
chemical_list: D010940:Plant Proteins; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases
keywords: Characterization; Glyceraldehyde-3-phosphate dehydrogenase; NAD(+) specific; Pea seed; Purification