Abstract
The bacterial phosphoenolpyruvate:glycose phosphotransferase system (PTS) plays a central role in catabolizing many sugars; regulation is effected by phosphorylation of PTS proteins. In Escherichia coli, the phosphoryltransfer sequence for glucose uptake is: PEP --> Enzyme I(His191) --> HPr(His15) --> IIIGlc(His90) --> IIGlc(Cys421) --> glucose. A rapid quench method has now been developed for determining the rate and equilibrium constants of these reactions. The method was validated by control experiments, and gave the following results for phosphoryltransfer between the following protein pairs. For phospho-HPr/IIIGlc (and HPr/phospho-IIIGlc), k1 = 6.1 x 10(7) M-1 s-1, k-1 = 4.7 x 10(7); for the mutant H75QIIIGlc in place of IIIGlc, k1 = 2.8 x 10(5) M-1 s-1, k-1 = 2.3 x 10(5). The derived Keq values agreed with the Keq obtained without use of the rapid quench apparatus. Keq for both reactions is 1-1.5. The rate of phosphoryltransfer between HPr and wild type IIIGlc is close to a diffusion-controlled process, while the reactions involving the mutant H75QIIIGlc are 200-fold slower. These rate differences are explained by an hypothesis for the mechanism of phosphoryltransfer between HPr and IIIGlc based on the structures of mutan...Continue Reading
References
Jun 15, 1978·Analytical Biochemistry·M A MarkwellN E Tolbert
Jun 1, 1976·Analytical Biochemistry·J P FroehlichR L Berger
May 1, 1979·Analytical Biochemistry·E B WaygoodS Roseman
Mar 11, 1992·Journal of Bacteriology·M H Saier, J Reizer
Apr 1, 1991·Protein Expression and Purification·P ReddyA Peterkofsky
Oct 15, 1991·Biochemistry·J G PeltonS Roseman
Jan 1, 1990·Annual Review of Biochemistry·N D MeadowS Roseman
Jun 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·K A PresperS Roseman
Jul 1, 1988·Journal of Bacteriology·D E ShevellG C Walker
Jan 1, 1987·Methods in Enzymology·T A KunkelR A Zakour
Nov 19, 1985·Biochemistry·E B WaygoodL T Delbaere
Jan 1, 1983·Canadian Journal of Biochemistry and Cell Biology = Revue Canadienne De Biochimie Et Biologie Cellulaire·R L Mattoo, E B Waygood
Oct 4, 1983·Biochimica Et Biophysica Acta·F F RoossienG T Robillard
Sep 1, 1994·Molecular Microbiology·M H Saier, J Reizer
Jan 1, 1994·Methods in Enzymology·C Frieden
Sep 1, 1993·Microbiological Reviews·P W PostmaG R Jacobson
Dec 27, 1996·The Journal of Biological Chemistry·J G PeltonS Roseman
Oct 1, 1964·Proceedings of the National Academy of Sciences of the United States of America·W KUNDIGS ROSEMAN
Citations
Jul 7, 1999·Biochimica Et Biophysica Acta·G T Robillard, J Broos
Nov 21, 1998·Molecular Microbiology·B M HogemaP W Postma
Jun 8, 2000·The Journal of Biological Chemistry·M G Gunnewijk, B Poolman
Apr 10, 1999·Molecular Biology of the Cell·R LuxS Khan
Nov 4, 2000·The EMBO Journal·G WangG M Clore
Dec 13, 2006·Microbiology and Molecular Biology Reviews : MMBR·Josef DeutscherPieter W Postma
Aug 25, 2016·PLoS Biology·Rahul SomavanshiVictor Sourjik
Jul 13, 2000·The Journal of Biological Chemistry·J M RohwerP W Postma
Dec 27, 1996·The Journal of Biological Chemistry·J G PeltonS Roseman
Jan 28, 2006·The Journal of Biological Chemistry·Norman D MeadowSaul Roseman
Mar 21, 2006·The Journal of Biological Chemistry·Himatkumar V PatelSaul Roseman
Oct 6, 2005·The Journal of Biological Chemistry·Norman D MeadowSaul Roseman
Jan 12, 1999·Journal of Bacteriology·M G GunnewijkB Poolman
Jan 24, 1998·Biochemistry·M D FeeseS J Remington
May 16, 1998·Biochemistry·D W PettigrewS J Remington