1998 Fiscal Year Final Research Report Summary
Structure and function of a new endothelial growth factor from snake venom
| Project/Area Number |
09672262
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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 | Meiji University |
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Principal Investigator |
KOMORI Yumiko Meiji University, Faculty of Pharmacy, Assistant Professor, 薬学部, 講師 (60162070)
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| Project Period (FY) |
1997 – 1998
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| Keywords | growth factor / endothelial cell / venom protein |
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| Research Abstract |
The heparin-binding dimeric hypotensive factor (HF) was purified from Vipera aspis aspis (Aspic viper) venom [Komori, Y. and Sugihara, H. (1990) Toxicon 28, 359-369]. In this study, the amino acid sequence, and structure and function of HF were elucidated. By electrospray ionization mass spectrometry (ESI-MS), the molecular weight of HF was determined to be 25,072.1. The complete amino acid sequence of HF was determined by Edman sequencing of the S-pyridylethylated HF and its peptides derived from enzymatic digestion. The theoretical molecular mass calculated from the primary structure agrees well with the molecular weight determined by ESI-MS. HF consists of two homogenous monomers bound covalently. The monomer with an N-terminal blocked by pyro-glutamic acid contains 110 amino acid residues including 8 cysteine residues, 2 of which are considered to be involved in intermolecular disulfide bonds. Sequential homology search revealed that the primary structure of HF is similar to that of vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) with a sequential homology of 45% and 22%, respectively. When injected intradermally into rat, an increase in capillary permeability was observed with HF or VEGF. On the other hand, only HF exerted a strong hypotensive effect immediately after intravenous injection into rat. Purified HF has a mitogenic effect on endothelial cells. Using bovine aortic endothelial cells (BAEC), the half maximal mitogenic concentration of HF was determined to be 5-5.5 nM (125-138 ng/ml). Similarly, VEGF had a mitogenic concentration at 0.5-1 nM. When incubated with HF and cycloheximide or HF and heparin, the cell growth was inhibited, suggesting that the mechanism of action of HF is similar to that of VEGF.
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