Abstract

The central objective of this proposal is to develop an understanding of the physical basis for protein-protein interactions. Several systems are to be studied, including the interaction of the apoptosis adaptor protein RAIDD with the prodomain of an ICH-1 protease; the putative N-terminal domains of the SV40 T antigen that interact with retinoblastoma protein and Hsc70; and the interaction of the T-cell surface proteins hCD2 and HCD58. In addition, an attempt will be made to convert miniproteins with a pre-defined scaffold structure into ligands of target proteins by random mutagenesis using phage display technologies. The principle structural method will be NMR spectroscopy and the thermodynamic properties of the various systems will be defined using isothermal titration calorimetry and surface plasmon resonances, as required. These studies are proposed to characterize protein-protein interactions and to develop an understanding of the underlying principles guiding the formation of specific protein-protein interfaces.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM038608-12
Application #
6179528
Study Section
Molecular and Cellular Biophysics Study Section (BBCA)
Program Officer
Wehrle, Janna P
Project Start
1990-09-01
Project End
2003-06-30
Budget Start
2000-07-01
Budget End
2001-06-30
Support Year
12
Fiscal Year
2000
Total Cost
$191,942
Indirect Cost

  Institution

Name
Harvard University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
082359691
City
Boston
State
MA
Country
United States
Zip Code
02115

  Publications

Marintchev, Assen; Edmonds, Katherine A; Marintcheva, Boriana et al. (2009) Topology and regulation of the human eIF4A/4G/4H helicase complex in translation initiation. Cell 136:447-60
Roehrl, Michael H A; Wang, Julia Y; Wagner, Gerhard (2004) A general framework for development and data analysis of competitive high-throughput screens for small-molecule inhibitors of protein-protein interactions by fluorescence polarization. Biochemistry 43:16056-66
Roehrl, Michael H A; Kang, Sunghyun; Aramburu, Jose et al. (2004) Selective inhibition of calcineurin-NFAT signaling by blocking protein-protein interaction with small organic molecules. Proc Natl Acad Sci U S A 101:7554-9
Roehrl, Michael H A; Wang, Julia Y; Wagner, Gerhard (2004) Discovery of small-molecule inhibitors of the NFAT--calcineurin interaction by competitive high-throughput fluorescence polarization screening. Biochemistry 43:16067-75
Roehrl, Michael H A; Hyberts, Sven G; Sun, Zhen-Yu Jim et al. (2003) Rapid backbone 1H, 13C, and 15N assignment of the V1 domain of human PKC iota using the new program IBIS. J Biomol NMR 26:373-4
Gross, John D; Gelev, Vladimir M; Wagner, Gerhard (2003) A sensitive and robust method for obtaining intermolecular NOEs between side chains in large protein complexes. J Biomol NMR 25:235-42
Lugovskoy, Alexey A; Degterev, Alexei I; Fahmy, Amr F et al. (2002) A novel approach for characterizing protein ligand complexes: molecular basis for specificity of small-molecule Bcl-2 inhibitors. J Am Chem Soc 124:1234-40
Muller, Jens; Lugovskoy, Alexey A; Wagner, Gerhard et al. (2002) NMR structure of the [2Fe-2S] ferredoxin domain from soluble methane monooxygenase reductase and interaction with its hydroxylase. Biochemistry 41:42-51
Zhou, P; Lugovskoy, A A; McCarty, J S et al. (2001) Solution structure of DFF40 and DFF45 N-terminal domain complex and mutual chaperone activity of DFF40 and DFF45. Proc Natl Acad Sci U S A 98:6051-5
Krezel, A M; Ulmer, J S; Wagner, G et al. (2000) Recombinant decorsin: dynamics of the RGD recognition site. Protein Sci 9:1428-38

Showing the most recent 10 out of 43 publications