2001 Fiscal Year Final Research Report Summary
The quantification of ratios between apo to holo types of metal binding protein : a new indicator of the oxidative stress in cells
| Project/Area Number |
11672314
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Laboratory medicine
|
|---|
| Research Institution | Osaka Medical College |
|---|
Principal Investigator |
NAKANISHI Toyofumi Osaka Medical College, Faculty of Medicine Assistant Professor, 医学部, 講師 (10247843)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
MIYAZAKI Ayako Osaka Medical College, Faculty of Medicine Research Associate, 医学部, 助手 (20298772)
KISHIKAWA Masahiko Osaka Medical College, Faculty of Medicine Research Associate, 医学部, 助手 (40268199)
SHIMIZU Akira Osaka Medical College, Faculty of Medicine Professor, 医学部, 教授 (00028581)
|
|---|
| Project Period (FY) |
1999 – 2001
|
| Keywords | NanoESIMS method / Ration of Apo / Holo / Oxidative stress in the body / Cooper-binding protein / Active oxide species / Free cooper ion / Neurodegenerative disease |
|---|
| Research Abstract |
We applied nano-electrospray ionization mass spectrometry (NanoESIMS) for the quantification of the rations between apo to holo types of metalloproteins (copper/zinc binding superoxide dismutase:SOD-1, metallothioneine) to clarify the etiology of neurodegenerative diseases such as familial amyotrophic lateral sclerosis. We examined the optimal conditions of ultra-sensitive mass spectrometric quantification of the ratios using metalloproteins standard and purified metalloproteins.
1) We examined the condition of pH range of sample solutions, contents of organic solvent and temperature of heated capillary.
2) We applied inductively coupled plasma mass spectrometry (ICPMS) instead of nanoESIMS technique.
Using the commercial SOD-1 standard, we could not detect dimer, holo type even though we modified some analytical conditions. We give up the construction of the optimal condition of NanoESIMS technique. On the other hand, we applied the ICPMS technique or the de e turn of holo type of SOD-1 and succeeded holo type of the metalloprotem. This technique (ICPMS) was a powerful tool of the quantification of the ratios between apo to holo types of metalloproteins.
|
