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. 1986 Jan 15;233(2):465–469. doi: 10.1042/bj2330465

Identification of an essential glutamic acid residue in beta-lactamase II from Bacillus cereus.

C Little, E L Emanuel, J Gagnon, S G Waley
PMCID: PMC1153048  PMID: 2869754

Abstract

Beta-Lactamase II from Bacillus cereus was readily inactivated by incubation at pH 4.75 with a water-soluble carbodiimide plus a suitable nucleophile. In the early stages of the reaction, 1 equivalent of nucleophile was incorporated/equivalent of enzyme, whereas during the later stages a second equivalent of nucleophile was also incorporated. This latter process correlated with the blocking of the enzyme's single thiol group. Enzyme inactivated in the presence of the coloured nucleophile N-(2,4-dinitrophenyl)ethylenediamine was fragmented by pepsin digestion, and coloured peptides were isolated by gel filtration and h.p.l.c. Two major peptides, representing 52% of the incorporated label, were isolated and sequenced. Both peptides contained the incorporated label on glutamic acid-37, and it is concluded that this latter residue represents a catalytically essential carboxylic residue in beta-lactamase II.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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