2002 Fiscal Year Final Research Report Summary
The role of proteins belonging to a plastin gene family in host defenses against bacterial infections.
| Project/Area Number |
13670275
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Bacteriology (including Mycology)
|
|---|
| Research Institution | Ehime University |
|---|
Principal Investigator |
SHINOMIYA Hiroto Ehime University School of Medicine, Dept. of Immunology and Host Defenses, Associate Professor, 医学部, 助教授 (80162618)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
ASANO Yoshihiro Ehime University School of Medicine, Dept. of Immunology and Host Defenses, Professor, 医学部, 教授 (70114353)
|
|---|
| Project Period (FY) |
2001 – 2002
|
| Keywords | p65 / L-plastin / phosphorylation / host defense / cytoskeleton / cell adhesion / actin / recombinant protein / immunostaining |
|---|
| Research Abstract |
We previously identified a 65-kDa protein (p65) that was phosphorylated in activated macrophages. It has turned out to be a murine homologue of human L-plastin that was identified as a novel protein in human cancer cells. P65/L-plastin is characterized by a series of Ca^<2+>-, calmodulin- and actin-binding domains, and is thought to play a crucial role in leukocytes and cancer cells. We have expressed a recombinant γ p65/L-plastin in E. coli that binds to b-actin and prepared high-titer antibodies using large amounts of the protein as immunogen. Anti-rp65/L-plastin antibodies recognize native p65/L-plastin as well as rp65/L-plastin and have enabled us to detect fine structures of intracellular p65/L-plastin, and it was found that its localization was extensively changed by stimulation with bacterial components. We further developed an enzyme-linked immunosorbent assay system and a flow cytometry method using these reagents, which made it possible to measure antibodies including autoantibodies against p65/L-plastin and to evaluate the maturation-dependent expression of the protein in leukocytes.
|
