| Project/Area Number |
17510177
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Living organism molecular science
|
|---|
| Research Institution | Mie University |
|---|
Principal Investigator |
INAGAKI Minoru Mie University, Department of Life Acicence, Associate Professor (20242935)
|
|---|
| Project Period (FY) |
2005 – 2007
|
| Project Status |
Completed (Fiscal Year 2007)
|
| Budget Amount *help |
¥2,150,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥150,000)
Fiscal Year 2007: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2006: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2005: ¥1,000,000 (Direct Cost: ¥1,000,000)
|
|---|
| Keywords | bacteriophage φX174 / spike H protein / lipoplvsaccharide / VS protease / protease resistant domain / surface plasmon resonance / conformational change / bacteriophageΦX174 / spikeHprotein / tryptophane residue / lipopolysaccharide / interaction / limited chemical degradation / protease-resistant domain / infection |
|---|
| Research Abstract |
Contribution of lipid part of LPS to the recognition by spike H protein
The LPS of E. coli C was treated with hydradine and potassium hydroxide to gave the deacylated derivatives of LPS. The derivatives showed significantly less affinity and small magnitude of conformational change toward the spike II protein of bacteriophage φX174 Contribution of charged residues of LPS to the recognition by spike H and G proteins
The derivatives of dephosphorylated and KDO reduced derivatives were prepared by the limited chemical degradation of LPS by hydrogen fluoride and sodium borohydride coupled with water soluble carbodiimide. These derivatives were submitted to the estimation of the interaction with the spike H and G proteins. The H protein was affected by the loss of phosphate residue and the G protein was affected by the loss of KDO residue from the LPS for the recognition.
Isolation and characterization of the protease resistance domains of spike H protein
By a mild treatment with VS protease, H protein gave two protease resistant domains(F2 and F3). The observations of N-terminal amino acid and MALDI-TOF-MS analyses revealed that. F2 corresponded to the fragment of 215-345 residues, on the other hand, F3 corresponded to the fragment of 1-102 residues of H protein. The fragment F2 showed strong affinity toward the LPS of host bacteria compared to the intact H protein, however, the fragment Fl showed 1/10 affinity to the intact H protein.
|
