title: Chicken liver glutamate dehydrogenase (GDH) demonstrates a histone H3 specific protease (H3ase) activity in vitro.
abstract: Site-specific proteolysis of the N or C-terminus of histone tails has emerged as a novel form of irreversible post-translational modifications assigned to histones. Though there are many reports describing histone specific proteolysis, there are very few studies on purification of a histone specific protease. Here, we demonstrate a histone H3 specific protease (H3ase) activity in chicken liver nuclear extract. H3ase was purified to homogeneity and identified as glutamate dehydrogenase (GDH) by sequencing. A series of biochemical experiments further confirmed that the H3ase activity was due to GDH. The H3ase clipped histone H3 products were sequenced by N-terminal sequencing and the precise clipping sites of H3ase were mapped. H3ase activity was only specific to chicken liver as it was not demonstrated in other tissues like heart, muscle and brain of chicken. We assign a novel serine like protease activity to GDH which is specific to histone H3.
authors.affiliation: Laboratory for Chromatin Biology, Department of Zoology, University of Delhi, North Campus, Delhi 110007, India. Electronic address: sachin.jogesh@gmail.com.
authors.forename: Jogeswar S
authors.initials: JS
authors.lastname: Purohit
authors.forename: Raghuvir S
authors.initials: RS
authors.lastname: Tomar
authors.forename: Anil K
authors.initials: AK
authors.lastname: Panigrahi
authors.forename: Shashibhal M
authors.initials: SM
authors.lastname: Pandey
authors.forename: Divya
authors.initials: D
authors.lastname: Singh
authors.forename: Madan M
authors.initials: MM
authors.lastname: Chaturvedi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002645:Chickens / Q000235:genetics; D010450:Endopeptidases / Q000737:chemistry / Q000378:metabolism; D005969:Glutamate Dehydrogenase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D008099:Liver / Q000201:enzymology*; D059748:Proteolysis
chemical_list: D006657:Histones; D005969:Glutamate Dehydrogenase; D010450:Endopeptidases; C021361:histone proteases
keywords: BNE; BSA; CNE; DTT; EDTA; EGTA; GAPDH; GDH; Glutamate dehydrogenase (GDH); H3ase; HMG; Histone H3 proteolysis; Histone H3-specific protease; IPTG; Irreversible modification of histone H3; LB; Luria–Bertani; MNase; Moonlighting protease; NLS; PCR; PMSF; bovine serum albumin; brain nuclear extract; chicken liver nuclear extract; dithiothreitol; ethylene glycol tetraacetic acid; ethylenediaminetetraacetic acid; glutamate dehydrogenase; glyceraldehyde-3-phosphate dehydrogenase; high mobility group; isopropyl β-d-1-thiogalactopyranoside; micrococcal nuclease; nuclear localization signal; phenylmethanesulfonylfluoride; polymerase chain reaction