Histidine-containing protein (HPr) is a phosphotransferase in the bacterial phosphoenolpyruvate-dependent phosphotransferase system. HPr provides a rich model system in which to study a number of important aspects of protein structure/function, including protein phosphorylation and protein-protein interactions. HPr can be phosphorylated at two different sites: phosphorylation of a histidine residue occurs as an intermediate in the phosphotransfer reaction and phosphorylation of a serine residue modulates the activity of the protein. The structural and dynamic ramifications of protein phosphorylation will be studied using HPr. As well, the interaction between HPr and its phosphoryl-acceptor protein, Enzyme IIA will be studied. To gain detailed information about these important aspects, solution structures of native HPr, its two phosphorylated forms, and mutant forms of the protein with altered functional properties. Continuing developments in NMR spectroscopy make this a powerful technique with which to probe the details of both structural and dynamical properties of proteins in solution.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK035187-10
Application #
2139518
Study Section
Molecular and Cellular Biophysics Study Section (BBCA)
Project Start
1985-04-01
Project End
1999-03-31
Budget Start
1995-04-01
Budget End
1996-03-31
Support Year
10
Fiscal Year
1995
Total Cost
Indirect Cost
Name
University of Washington
Department
Biochemistry
Type
Schools of Medicine
DUNS #
135646524
City
Seattle
State
WA
Country
United States
Zip Code
98195
Lee, S Y; Klevit, R E (2000) The whole is not the simple sum of its parts in calmodulin from S. cerevisiae. Biochemistry 39:4225-30
Rohl, C A; Boeckman, F A; Baker, C et al. (1999) Solution structure of the sodium channel inactivation gate. Biochemistry 38:855-61
Rajagopal, P; Jones, B E; Klevit, R E (1998) Solvent exchange rates of side-chain amide protons in proteins. J Biomol NMR 11:205-12
Rajagopal, P; Waygood, E B; Reizer, J et al. (1997) Demonstration of protein-protein interaction specificity by NMR chemical shift mapping. Protein Sci 6:2624-7
Jones, B E; Rajagopal, P; Klevit, R E (1997) Phosphorylation on histidine is accompanied by localized structural changes in the phosphocarrier protein, HPr from Bacillus subtilis. Protein Sci 6:2107-19
Jones, B E; Dossonnet, V; Kuster, E et al. (1997) Binding of the catabolite repressor protein CcpA to its DNA target is regulated by phosphorylation of its corepressor HPr. J Biol Chem 272:26530-5
Thapar, R; Nicholson, E M; Rajagopal, P et al. (1996) Influence of N-cap mutations on the structure and stability of Escherichia coli HPr. Biochemistry 35:11268-77
Peterkofsky, A; Seok, Y J; Amin, N et al. (1995) The Escherichia coli adenylyl cyclase complex: requirement of PTS proteins for stimulation by nucleotides. Biochemistry 34:8950-9
Pullen, K; Rajagopal, P; Branchini, B R et al. (1995) Phosphorylation of serine-46 in HPr, a key regulatory protein in bacteria, results in stabilization of its solution structure. Protein Sci 4:2478-86
Herzberg, O; Klevit, R (1994) Unraveling a bacterial hexose transport pathway. Curr Opin Struct Biol 4:814-22

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