Glucosamine-6-phosphate synthase from Escherichia coli: Determination of the mechanism of inactivation by N sup 3 -fumaroyl-L-2,3-diaminopropionic derivatives
- Laboratoire de Bioorganique et Biotechnologies, Paris (France)
A mechanistic investigation of the inactivation of Escherichia coli glucosamine-6-phosphate synthase by N{sup 3}-(4-methoxyfumaroyl)-L-2,3-diaminopropionate (FMDP) was undertaken. On the basis of the known participation of the N-terminal cysteine residue in this process, the model reactions between FMDP and L-cysteine and between FMDP and the synthetic decapeptide Cys-Gly-Ile-Val-Gly-Ala-Ile-Ala-Ile-Ala-Gln-Arg, corresponding to the amino-terminal protein sequence, were studied. The results allowed us to propose a pathway that is in perfect agreement with the biochemical results: enzyme inactivation arose from Michael addition of glutamine binding site cysteine-1 on the fumaroyl double bond at the {beta}-position of the ester group. Upon denaturation under slightly alkaline conditions, this adduct underwent cyclization to a transient succinimide adduct, which rearranged into the stable 2-substituted 1,4-thiazin-3-one-5-carboxylate involving participation of the cysteine amino group. The tryptic radiolabeled peptides purified from ({sup 3}H)FMDP-treated enzyme and resistant to Edman degradation coeluted with the products resulting from the model reaction between the synthetic decapeptide and the inhibitor.
- OSTI ID:
- 6324622
- Journal Information:
- Biochemistry; (USA), Journal Name: Biochemistry; (USA) Vol. 29:15; ISSN 0006-2960; ISSN BICHA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
AMINES
BACTERIA
CARBOHYDRATES
CARBON 14 COMPOUNDS
CARBOXYLIC ACIDS
CHEMICAL REACTIONS
CHROMATOGRAPHY
DERIVATIZATION
ENZYMES
ESCHERICHIA COLI
GLUCOSAMINE
HEXOSAMINES
HEXOSES
INACTIVATION
LABELLED COMPOUNDS
LIQUID COLUMN CHROMATOGRAPHY
MICROORGANISMS
MONOCARBOXYLIC ACIDS
MONOSACCHARIDES
ORGANIC ACIDS
ORGANIC COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
PROPIONIC ACID
PURIFICATION
SACCHARIDES
SEPARATION PROCESSES
THIN-LAYER CHROMATOGRAPHY
TRANSFERASES