Interactive and dominant effects of residues 128 and 141 on cyclic nucleotide and DNA bindings in Escherichia coli cAMP receptor protein.

The Journal of Biological Chemistry
X Cheng, J C Lee

Abstract

The molecular events in the cAMP-induced allosteric activation of cAMP receptor protein (CRP) involve interfacial communications between subunits and domains. However, the roles of intersubunit and interdomain interactions in defining the selectivity of cAMP against other cyclic nucleotides and cooperativity in ligand binding are still not known. Natural occurring CRP mutants with different phenotypes were employed to address these issues. Thermodynamic analyses of subunit association, protein stability, and cAMP and DNA binding as well as conformational studies of the mutants and wild-type CRPs lead to an identification of the apparently dominant roles of residues 128 and 141 in the cAMP-modulated DNA binding activity of CRP. Serine 128 and the C-helix were implicated as playing a critical role in modulating negative cooperativity of cyclic nucleotide binding. A correlation was established between a weak affinity for subunit assembly and the relaxation of cyclic nucleotide selectivity in the G141Q and S128A/G141Q mutants. These results imply that intersubunit interaction is important for cyclic nucleotide discrimination in CRP. The double mutant S128A/G141Q, constructed from two single mutations of S128A and G141Q, which exhib...Continue Reading

References

Jan 1, 1979·Methods in Enzymology·J C Lee, S N Timasheff
Mar 1, 1992·Microbiological Reviews·J L Botsford, J G Harman
Oct 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·J KimS Garges
Mar 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·T Heyduk, J C Lee
Oct 1, 1988·Tissue Antigens·T D LeeD X Cheng
Jun 1, 1971·Proceedings of the National Academy of Sciences of the United States of America·A D RiggsG Zubay
Apr 1, 1973·Proceedings of the National Academy of Sciences of the United States of America·R Sanders, D McGeoch
Sep 1, 1973·Proceedings of the National Academy of Sciences of the United States of America·J S Krakow, I Pastan
Feb 25, 1982·Nucleic Acids Research·P Cossart, B Gicquel-Sanzey
Jan 25, 1984·Journal of Molecular Biology·M G Fried, D M Crothers
May 25, 1984·Science·B de CrombruggheH Buc
Dec 1, 1981·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·T O Sippel
Jan 1, 1993·Annual Review of Biochemistry·A KolbS Adhya
Mar 1, 1964·Biochemistry·D A YPHANTIS

❮ Previous
Next ❯

Citations

Mar 11, 2008·International Journal of Biological Macromolecules·Xianrong ShenShaoning Yu
Nov 16, 2002·The Journal of Biological Chemistry·Hwan YounGary P Roberts
May 22, 2001·The Journal of Biological Chemistry·M V ThorsteinssonG P Roberts

❮ 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.