2001 Fiscal Year Final Research Report Summary
Measurement of nitric oxide in animal tissues with monoclonal antibodies
Project/Area Number |
12660275
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Research Category |
Grant-in-Aid for Scientific Research (C)
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Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Basic veterinary science/Basic zootechnical science
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Research Institution | OSAKA PREFECTURE UNIVERSITY |
Principal Investigator |
SUYAMA Shingo Agriculture and Life Science, Osaka Prefecture University, Professor, 農学生命科学研究科, 教授 (00094508)
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Project Period (FY) |
2000 – 2001
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Keywords | Nitric-Oxide / Monoclonal Antibody / soluble guanylate cyclase / Deletion Mutant / Animal Tissues |
Research Abstract |
For examination of the interaction between monoclonal antibody (mAb3221) against NO-binding soluble guanylate cyclase and soluble guanylate cyclase purified from bovine lungs (sGC), a Fison lasys instrument that measures surface plasmon resonance was used. With mAb3221 and phosphate-buffered saline containing 0.1 % Triton X-100 (PBST) not treated with NO, Kd equaled 1.12 X 10^<-6>M for sGC. When PBST was treated with NO, Kd value for sGC was 1.22 X 10^<-8>M. When carbon monoxide (CO) was bubbled through the PBST, the Kd for sGC was 0.17 X 10^<-7> M. Although YC-1, an sGC activator did not affected Kd for sGC, the activity of this enzyme was increased. When YC-1 and CO were used to PBST, sGC activity increased 100-times, the Kd value was close to that with NO treatment. These compounds may interfere with the measurement of NO. In examination of insect cells with some kinds deletion mutant of sGC beta-subunit, Kd= 1.05 X 10^<-8>M for mAb3221 in the presence of NO. Therefore, NO can be measured with mAb3221 and the deletion mutant tested.
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Research Products
(5 results)