Characterization of disulfide exchange between DsbA and HtrA proteins from Escherichia coli.

Joanna Skórko-Glonek; Anna Sobiecka-Szkatuła; Barbara Lipińska

doi:10.18388/abp.2006_3331

Characterization of disulfide exchange between DsbA and HtrA proteins from Escherichia coli.

Joanna Skórko-Glonek Department of Biochemistry, University of Gdańsk, Gdańsk, Poland. glonek@biotech.univ.gda.pl;
Anna Sobiecka-Szkatuła
Barbara Lipińska

DOI: https://doi.org/10.18388/abp.2006_3331

Abstract

DsbA is the major oxidase responsible for generation of disulfide bonds in proteins of E. coli envelope. In the present work we provided the first detailed characterization of disulfide exchange between DsbA and its natural substrate, HtrA protease. We demonstrated that HtrA oxidation relies on DsbA, both in vivo and in vitro. We followed the disulfide exchange between these proteins spectrofluorimetrically and found that DsbA oxidizes HtrA with a 1:1 stoichiometry. The calculated second-order apparent rate constant (kapp) of this reaction was 3.3x10(4)+/-0.6x10(4) M-1s-1. This value was significantly higher than the values obtained for nonfunctional disulfide exchanges between DsbA and DsbC or DsbD and it was comparable to the kapp values calculated for in vitro oxidation of certain non-natural DsbA substrates of eukaryotic origin.

Published

2006-10-01

Issue

Vol. 53 No. 3 (2006)

Section

Articles

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