Properties of the high-affinity single-stranded DNA binding state of the Escherichia coli recA protein

Joseph P. Menetski, Abraham Varghese, Stephen C. Kowalczykowski

Research output: Contribution to journalArticlepeer-review

68 Scopus citations

Abstract

The properties of the high-affinity single-stranded DNA (ssDNA) binding state of Escherichia coli recA protein have been studied. We find that all of the nucleoside triphosphates that are hydrolyzed by recA protein induce a high-affinity ssDNA binding state. The effect of ATP binding to recA protein was partially separated from the ATP hydrolytic event by substituting calcium chloride for magnesium chloride in the binding buffer. Under these conditions, the rate of ATP hydrolysis is greatly inhibited. ATP increases the affinity of recA protein for ssDNA in a concentration-dependent manner in the presence of both calcium and magnesium chloride with apparent Kd values of 375 and 500 μ ATP, respectively. Under nonhydrolytic conditions, the molar ratio of ATP to ADP has an effect on the recA protein ssDNA binding affinity. Over an ATP/ADP molar ratio of 2-3, the affinity of recA protein for ssDNA shifts cooperatively from a low-to a high-affinity state.

Original languageEnglish (US)
Pages (from-to)1205-1212
Number of pages8
JournalBiochemistry
Volume27
Issue number4
StatePublished - 1988
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

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