2001 Fiscal Year Final Research Report Summary
Structure and function of three types of phospholipase A_2 inhibitor s derived from the blood of venomous snakes
| Project/Area Number |
12672137
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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 |
Biological pharmacy
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| Research Institution | Osaka University of pharmaceutical Sciences, Assistant |
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Principal Investigator |
INOUE Seiji Faculty of Pharmacy, Osaka University of Pharmaceutical Sciences Associate Profesor, 薬学部, 助教授 (70183184)
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| Project Period (FY) |
2000 – 2001
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| Keywords | Phospholipase A_2 / Inhibitor / Venomous snake / Serum protein / Tertiary structure / Crystallization / Structure function relationship / Inhibition mechanism |
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| Research Abstract |
1. Structure and function of PLIα
We have identified a PLIα-like protein, which showed about 70% sequence homology to Chinese mamushi PLIα but showed no iuhibitory activity from the serum of the non-venorilous striated snake and cloned its cDNA. We have constructed the recombinant chiineric proteins between the PLIα-like protein and Chinese mamushi PLIα and found that the sequence from 13 to 36 residues of Chinese mamushi PLIα were important for the bindihg and inhibition of the PLA_<2->. Furthermore, we found that the amino acid residues 26, 28, and 29 were especially important among this region. On the other hand, in order to determine the tertiary structure of PLIα, we searched for the best crystallization conditions of PLIα purified from the Chinese mamushi, serum, and we could obtain its needle crystal. However, the X-ray diffiaction image is not obtained yet.
2. Structure and function of PLIα
We have identified PLIβ, which specifically inhibit group-II basic protein, from the serum of the striated snake, purified it, and cloned its cDNA. Alternatively, we are tying to establish the E. coli expression system for Chinese mamushi PLIβ. On the other hand, we searched for the best crystallization conditions of PLIβ purified from Chinese mamushi, serum, but we could not obtain the crystals suitable for X-ray crystallography by this time
3. Structure and function of PLIγ
We are investigating whether PLIγ could be reconstructed from the separated two subunits of Chinese mamushi PLIγ Without a loss of the inhibitory activity. On the other hand, we searched, for the best crystallization conditions of PLIγ purified from Chinese mamushi, serum, but we could not obtain the crystals suitable for X-ray crystallography by this time.
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Research Products
(2 results)
