Substrate-decreased modification by diethyl pyrocarbonate of two histidines in isocitrate lyase from Escherichia coli

Journal Article · Tue Jul 30 04:00:00 EDT 1991 · Biochemistry; (United States)

DOI:https://doi.org/10.1021/bi00244a012· OSTI ID:5047469

Ko, Young Hee; Vanni, P; Munske, G R; McFadden, B A ^[1]

---

1. Washington State Univ., Pullman (United States)

The inactivation of tetrameric 188-kDa isocitrate lyase from Escherichia coli at pH 6.8 (37{degrees}C) by diethyl pyrocarbonate, exhibiting saturation kinetics, is accompanied by modification of histidine residues 266 and 206. Substrates isocitrate, glyoxylate, or glyoxylate plus succinate protect the enzyme from inactivation, but succinate alone does not. Removal of the carbethoxy groups from inactivated enzyme by treatment with hydroxylamine restores activity of isocitrate lyase. The present results suggest that the group-specific modifying reagent diethyl pyrocarboate may be generally useful in determining the position of active site histidine residues in enzymes.

• OSTI ID: 5047469
• Journal Information: Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 30:30; ISSN 0006-2960; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
ALDEHYDES
AMINO ACIDS
AZOLES
BACTERIA
BIOLOGICAL EFFECTS
CARBONIC ACID ESTERS
CARBOXYLIC ACID SALTS
CARBOXYLIC ACIDS
CITRATES
ENZYMES
ESCHERICHIA COLI
ESTERS
GLYOXYLIC ACID
HETEROCYCLIC ACIDS
HETEROCYCLIC COMPOUNDS
HISTIDINE
IMIDAZOLES
INACTIVATION
LYASES
MICROORGANISMS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
SUBSTRATES