| Project/Area Number |
11660292
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Basic veterinary science/Basic zootechnical science
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| Research Institution | Iwate University |
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Principal Investigator |
SYUTO Bunei Iwate University, Veterinary Medicine, Professor, 農学部, 教授 (60001533)
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| Co-Investigator(Kenkyū-buntansha) |
MORIMATSU Masami Iwate University, Veterinary Medicine, Associate Professor, 農学部, 助教授 (70241370)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2000: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1999: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | chitin binding protein / bivine / IgM / sugar chain / GlcNAc / serum proteins / opsonin / zymosan / 活性酸素 / N-アセチルグルコサミン / Nーアセチルグルコサミン |
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| Research Abstract |
The activation of the neutorophil function, such as phagocytosis and superoxide production induced by opsonins, is a very important system in the nonspecific defense, and its mechanism has been studied extensively. On the other hand, we have hypothesized that some kinds of regulatory proteins to the neutrophil activation system are present in serum and play the important roles as a regulatory systems. This hypothesis motivates us to isolate and to characterize a regulatory protein in the bovine serum. In this study, a bovine serum protein which binds to chitin (chitin binding protein, BPb01) and suppresses the superoxide generation in neutorphils has been purified and identified. The outline of the studies is as follows.
1) A protein with high affinity to N-acetyl-D-glucosamine (GlcNAc) was separated and was named CBPb01.
2) Purification and identification : the amino acid sequence of CBPb01 peptides were identical to the reported amino acid sequence of bovine IgM.
3) Constituent carbohydrates : lectin binding analysis revealed that CBPb01 contains mannose, GlcNAc and fucose.
4) Inhibition of superoxide generation by CBPb01 : about 40% of superoxide generation was decreased, when a neutrophils were stimulated with the opsonized zymosan that was treated with CBPb01. This decrease was completely inhibited by adding GlcNAc.
We have concluded that CBPb01 is not a lectin but a GlcNAc recognizing IgM that regulate the superoxide generation in neutrophils.
5) Inhibition mechanism for the superoxide generation by CBPb01 : although CBPb01 binds to neutrophils, the superoxide generation was not decreased. These results suggest that CBPbO1 inhibits the active structure of protein complex formed with opsonins, while it does not involve the process of the down stream from phospholipase C to NADPH oxidase activation in the superoxide generation system.
6) This is the first report of the chitin-binding bovine IgM that controls a superoxide generation in neutrophils.
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