title: Characterization of disulfide exchange between DsbA and HtrA proteins from Escherichia coli.
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.
authors.affiliation: Department of Biochemistry, University of Gdańsk, Gdańsk, Poland. glonek@biotech.univ.gda.pl
authors.forename: Joanna
authors.initials: J
authors.lastname: Skórko-Glonek
authors.forename: Anna
authors.initials: A
authors.lastname: Sobiecka-Szkatuła
authors.forename: Barbara
authors.initials: B
authors.lastname: Lipińska
mesh_terms: D004220:Disulfides / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000378:metabolism*; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000737:chemistry / Q000502:physiology; D019897:Periplasm / Q000201:enzymology* / Q000378:metabolism; D033903:Periplasmic Proteins / Q000737:chemistry / Q000378:metabolism*; D019704:Protein Disulfide-Isomerases / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins; D012697:Serine Endopeptidases / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D004220:Disulfides; D006360:Heat-Shock Proteins; D033903:Periplasmic Proteins; D011994:Recombinant Proteins; D010088:Oxidoreductases; C061682:DegP protease; D012697:Serine Endopeptidases; D019704:Protein Disulfide-Isomerases