2005 Fiscal Year Final Research Report Summary
Analysis of signaling network by protein-interaction proteomics
| Project/Area Number |
15201044
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (A)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Applied genomics
|
|---|
| Research Institution | The University of Tokushima |
|---|
Principal Investigator |
TANIGUCHI Hisaaki The University of Tokushima, The Institute for Enzyme Research, Professor, 分子酵素学研究センター, 教授 (10257636)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
FUJIWARA Kazuko The University of Tokushima, The Institute for Enzyme Research, Assistant professor, 分子酵素学研究センター, 助教授 (20108880)
KONISHI Hiroaki The University of Tokushima, The Institute for Enzyme Research, Assistant professor, 分子酵素学研究センター, 助教授 (40252811)
IKEDA Kazuko The University of Tokushima, The Institute for Enzyme Research, Assistant, 分子酵素学研究センター, 助手 (10108863)
YAMAUCHI Emiko The University of Tokushima, The Institute for Enzyme Research, Assistant, 分子酵素学研究センター, 助手 (50332292)
MATSUZAKI Hideki RIKEN, Posttranslational Modification and Dynamic Regulation Research, Team Researcher, 翻訳後修飾による動的調節機構研究チーム, 連携研究員 (30392129)
|
|---|
| Project Period (FY) |
2003 – 2005
|
| Keywords | phosphorylation / proteomics / signaling network / protein-interaction |
|---|
| Research Abstract |
To understand molecular mechanisms underlying protein modification-dependent protein-protein interactions, protein crystals of MARCKS, a major C kinase substrate protein, were produced as calmodulin complex. Another target was calmodulin complexed with the N-terminal myristoylated domain of neuron specific NAP-22 protein. The X-ray structure of the former complex revealed the phosphorylation-dependent protein-protein interaction involved in the MARCKS-calmodulin interaction. The latter structure demonstrated that the Myristoyl moiety in directly involved in the NAP-22-calmodulin interaction. The second research project deals with the proteomic analysis of cellular organelles. An antibody-based purification method was established to obtain ‘pure' peroxisomes from rat liver. Protein components including several novel proteins were elucidated. A novel peroxisome-specific Lon protease, a molecular chaperon, was discovered. Another organelle, lipid droplet, was analyzed similarly, and one of small G proteins, Rab 18, was found to be localized in the organelle. The third research project was to analyze signaling networks that involve protein phosphorylation and protein interaction networks. As a model system, the EGF receptor signaling networks were analyzed by isolating tyrosine-phosphorylated proteins by phospho-proteome analysis. About 150 proteins that were either tyrosine phosphorylated themselves or proteins interacting with phosphorylated proteins were identified. About one third of them were novel proteins. The physiological functions of these proteins were elucidated by analyzing interacting proteins and EGF-dependent phosphorylation. Several of them were found to be involved in the regulation of EGF receptor downregulation. Antibodies that recognize these proteins and several phosphorylation sites were raised and used to analyze the signaling networks downstream of the EGF receptor.
|
Research Products
(25 results)
