Structures of active site histidine mutants of IIIGlc, a major signal-transducing protein in Escherichia coli. Effects on the mechanisms of regulation and phosphoryl transfer.

The Journal of Biological Chemistry
J G PeltonS Roseman

Abstract

IIIGlc (also called IIAGlc), a major signal-transducing protein in Escherichia coli, is also a phosphorylcarrier in glucose uptake. The crystal and NMR structures of IIIGlc show that His90, the phosphoryl acceptor, adjoins His75 in the active site. Glutamine was substituted for His-, giving H75QIIIGlc and H90QIIIGlc, respectively (Presper, K. A., Wong, C.-Y., Liu, L., Meadow, N. D., and Roseman, S. (1989) Proc. Natl. Acad. Sci. U. S. A. 86, 4052-4055), but the mutants showed unexpected properties. H90QIIIGlc loses regulatory functions of IIIGlc, and the phosphoryltransfer rates between HPr/H75QIIIGlc are 200-fold less than HPr/IIIGlc (Meadow, N. D., and Roseman, S. (1996) J. Biol. Chem. 271, 33440-33445). X-ray crystallography, differential scanning calorimetry, and NMR have now been used to determine the structures of the mutants (phospho-H75QIIIGlc was studied by NMR). The three methods gave completely consistent results. Except for the His to Gln substitutions, the only significant structural changes were in a few hydrogen bonds. H90QIIIGlc contains two structured water molecules (to Gln90), which could explain its inability to regulate glycerol kinase. Phospho-IIIGlc contains a chymotrypsin-like, hydrogen bond network (Thr7...Continue Reading

References

May 1, 1979·Analytical Biochemistry·E B WaygoodS Roseman
Dec 7, 1977·Journal of the American Chemical Society·F BlombergH Rüterjans
Nov 1, 1992·Acta Crystallographica. Section A, Foundations of Crystallography·D E Tronrud
Oct 30, 1991·Journal of Molecular Biology·G KapadiaO Herzberg
Dec 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·D WorthylakeS J Remington
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
Jan 1, 1985·Methods in Enzymology·A J HowardN H Xuong
Sep 1, 1974·Journal of Bacteriology·F C NeidhardtD F Smith
Jan 1, 1984·Progress in Biophysics and Molecular Biology·E N Baker, R E Hubbard
Sep 1, 1994·Molecular Microbiology·M H Saier, J Reizer
Dec 1, 1994·American Journal of Respiratory Cell and Molecular Biology·D MichelisJ R Wright
Apr 26, 1994·Proceedings of the National Academy of Sciences of the United States of America·M FeeseS J Remington
Sep 1, 1993·Microbiological Reviews·P W PostmaG R Jacobson
Jul 1, 1996·Protein Science : a Publication of the Protein Society·S M Habermann, K P Murphy
Dec 9, 1994·Science·C L Perrin

❮ Previous
Next ❯

Citations

Oct 29, 1998·Protein Science : a Publication of the Protein Society·R L van Montfort, B W Dijkstra
Jul 26, 2008·Biochemistry·Jared B Parker, James T Stivers
Mar 21, 2006·The Journal of Biological Chemistry·Himatkumar V PatelSaul Roseman
Jan 26, 2006·Journal of the American Chemical Society·Jianhua XuJay R Knutson

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.