2005 Fiscal Year Final Research Report Summary
Development of NMR strategies for rapid structure determination of protein complexes
| Project/Area Number |
16570090
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Structural biochemistry
|
|---|
| Research Institution | HOKKAIDO UNIVERSITY |
|---|
Principal Investigator |
OGURA Kenji Hokkaido Univ., Grad.School of Pharm.Sci., Instructor, 大学院・薬学研究科, 助手 (50270682)
|
|---|
| Project Period (FY) |
2004 – 2005
|
| Keywords | NMR / protein / structure |
|---|
| Research Abstract |
The tetratricopeptide repeat (TPR) domain of p67phox, which is a part of NADPH oxidase, plays an important role in activation of the enzyme by binding the Rac protein. I and co-workers accompolished the sequence- and methyl-specific resonance assignment of TPR domain of p67phox using uniformly 2H/13C/15N-labeled protein containing protonated methyl moieties of Val, Leu and Ile residues. Furthermore, the global foldings of the protein were determined using NMR spectroscopy. This strategy could be applied to protein-protein complexes.
The solution structure of growth factor receptor-bound protein 2 (Grb2) SH2 complexed with a high-affinity inhibitor, having a non phosphorus moiety and a macrocyclic peptide mimetic, was determined by nuclear magnetic resonance (NMR) spectroscopy. In the process of the investigation, I prepared the NMR-sample of molecular complex of the inhibitor and 2H/15N-labeled Grb2 SH2. Deutration of the protein led to simplification of NMR signals. The solution structure showed that overall conformation of the inhibitor on the molecular surface of Grb2 SH2 was very similar to that of phosphotyrosine-containing ligand peptide derived from BCR-Abl. The binding interaction between the inhibitor and Grb2 SH2 in the solution structure proved that the structure-based drug design was appropriate. Based on the structure, it will be possible to design new generation inhibitors with similar binding mechanism.
|
