Overexpression and sequence of the Escherichia coli cheY gene and biochemical activities of the CheY protein.
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Matsumura P, Rydel JJ, Linzmeier R, Vacante D
Overexpression and sequence of the Escherichia coli cheY gene and biochemical activities of the CheY protein.
J Bacteriol. 1984 Oct;160(1):36-41.
- PubMed ID
- 6090423 [ View in PubMed]
- Abstract
We overexpressed the CheY protein by fusing the cheY gene to the tryptophan promoter from Serratia marcescens. Expression of the trp promoter-cheY fusion and subsequent purification of the protein resulted in the isolation of up to 20 mg of homogeneously pure CheY protein from 100 mg of the cytoplasmic supernatant fraction. Purification of the CheY protein was accomplished by exploiting the affinity of CheY protein to cibacron blue dye and molecular sieve chromatography. Preliminary biochemical characterization of the pure CheY protein revealed specific interactions with S-adenosylmethionine and cibacron blue dye. Additional kinetic analysis showed that CheY protein inhibits EcoRI methyltransferase. The amino acid composition of the CheY protein predicted by the DNA sequence of the cheY gene and the amino acid analysis of the CheY protein were in agreement, confirming the authenticity of the purified CheY protein.