Synthesis of 1-chloro-2-oxohexanol 6-phosphate, a covalent active-site reagent for phosphoglucose isomerase

Klaus D. Schnackerz, John Chirgwin, Ernst A. Noltmann

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

A new covalent active site reagent, 1-chloro-2-oxo-6-hexanol 6-phosphate, has been synthesized from glutaric acid monomethyl ester and characterized by NMR spectroscopy. Inactivation of phosphoglucose isomerase, when incubated with various modifier concentrations, was found to be pseudo first order with respect to enzyme concentration (half-life of inactivation 6 h at pH 7.5 (30°C) and 2.0 μM active site concentration) but showed saturation kinetics for the dependence on inactivator concentration. This saturation phenomenon demonstrates the occurrence of a reversible enzyme-inhibitor complex (Kdiss = 14.3 mM) preceding the irreversible inactivation via the chloromethyl oxo groups. Substrate or competitive inhibitors such as 6-phosphogluconate or 5-phosphoarabinonate protect against inactivation of the isomerase by the modifying reagent.

Original languageEnglish (US)
Pages (from-to)1756-1761
Number of pages6
JournalBiochemistry
Volume20
Issue number7
StatePublished - 1981
Externally publishedYes

Fingerprint

Glucose-6-Phosphate Isomerase
Catalytic Domain
Hexanols
Isomerases
Enzyme Inhibitors
Nuclear magnetic resonance spectroscopy
Half-Life
Esters
Magnetic Resonance Spectroscopy
Phosphates
Kinetics
Substrates
Enzymes
1-chloro-2-ketohexanol-6-phosphate
6-phosphogluconic acid
glutaric acid
arabinonate-5-phosphate

ASJC Scopus subject areas

  • Biochemistry

Cite this

Synthesis of 1-chloro-2-oxohexanol 6-phosphate, a covalent active-site reagent for phosphoglucose isomerase. / Schnackerz, Klaus D.; Chirgwin, John; Noltmann, Ernst A.

In: Biochemistry, Vol. 20, No. 7, 1981, p. 1756-1761.

Research output: Contribution to journalArticle

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