Expression and site-directed mutagenesis of hepatic glucokinase

A. J. Lange, L. Z. Xu, F. Van Poelwijk, K. Lin, D. K. Granner, S. J. Pilkis

Research output: Contribution to journalArticlepeer-review

28 Scopus citations

Abstract

Soluble rat liver glucokinase was expressed at high levels at 22°C in the BL21(DE3)pLysS strain of Escherichia coli. Aspartate-211 of yeast hexokinase has been implicated as a catalytic residue from crystallographic data. The corresponding residue in rat liver glucokinase, aspartate-205, was mutated to alanine and the expressed mutant had 1/500th of the activity of the wild type, with no change in the K(m) values for glucose or ATP. The results support a role for this residue as a base catalyst in the glucokinase reaction and, most probably, a similar role in the reactions of all members of the hexokinase family.

Original languageEnglish (US)
Pages (from-to)159-163
Number of pages5
JournalBiochemical Journal
Volume277
Issue number1
DOIs
StatePublished - 1991

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