title: Kinetics and thioredoxin specificity of thiol modulation of the chloroplast H+-ATPase.
abstract: The kinetics of thiol modulation of the chloroplast H+-ATPase (CF0CF1) in membrana were analyzed by employing thioredoxins that were kept reduced by 0.1 mM dithiothreitol. The kinetics of thiol modulation depend on the extent of the proton gradient. The process is an exponential function of the thioredoxin concentration and reaction time and can be described by an irreversible second order reaction. The results indicate that the formation of the complex between thioredoxin and CF0CF1 is slow compared with the subsequent reduction step. Furthermore we have compared the efficiencies of the Escherichia coli thioredoxin Trx and the two chloroplast thioredoxins Tr-m and Tr-f. The second order rate constants are 0.057 (Tr-f), 0.024 (Trx), and 0.010 s-1 microM-1 (Tr-m) suggesting that Tr-f rather than Tr-m is the physiological reductant for the chloroplast ATPase. The often employed artificial reductant dithiothreitol exhibits a second order rate constant in thiol modulation of 1.02.10(-6) s-1 microM-1.
authors.affiliation: Institut für Biochemie der Pflanzen, Heinrich-Heine-Universität Düsseldorf, Universitätsstrasse 1, D-40225 Düsseldorf, Federal Republic of Germany.
authors.forename: O
authors.initials: O
authors.lastname: Schwarz
authors.forename: P
authors.initials: P
authors.lastname: Schürmann
authors.forename: H
authors.initials: H
authors.lastname: Strotmann
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D004926:Escherichia coli; D005998:Glycine / Q000031:analogs & derivatives / Q000378:metabolism; D006868:Hydrolysis; D007700:Kinetics; D006180:Proton-Translocating ATPases / Q000378:metabolism*; D018724:Spinacia oleracea; D013438:Sulfhydryl Compounds / Q000378:metabolism*; D013879:Thioredoxins / Q000378:metabolism*; D013997:Time Factors
chemical_list: D013438:Sulfhydryl Compounds; D013879:Thioredoxins; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases; D005998:Glycine; C100184:tricine