강원도 지역에 서식하는 무당개구리에서 생물학적 활성을 지닌 Bombesin 유사 Peptide 에 관한 연구 = Studies on Biologically active bombesin - like Peptides in frog ( Bombina orientalis ) inhabiting Kangwon - Do

다국어 초록 (Multilingual Abstract)

The present study was performed to purify a BLI from the skin of frogs, B. orientalis inhabiting Kangwon-Do, Korea, and then determine its physiological activities and amino acid sequence. A substance that shows bombesin-like immunoreactivity (BLI) was partially purified from crude methanol extract of the frog skin by using a column of alkaline alumina and Sephadex G-10. BLI was further purified by using RP C18 preparative HPLC in which BLI was separated into 5 peaks. The major peak (BLI-K1) containing 65% of total BLI was subjected to purify as a single component by using sequential HPLC of SP-ion exchange and RP C18. Content of BLI in each fraction was monitored by RIA for which bombesin antiserum with a high titer and affinity to BBS was raised in a guinea pig. Eventually, 890 ㎍ of BLI-K1 was homogeneously purified from 420g of the skin of B. orientalis and it was not differentiated from synthetic BBS in RP C18, gel permeation and SP-ion exchange HPLC. In the physiological activities, BLI-K1 was identical to synthetic BBS in stimulation of gastrin release and exocrine pancreatic secretion including volume, protein, bicarbonate, amylase and chymotrypsin output in anesthetized rats. Furthermore, molecular weight and partial amino acid sequence of BLI-K1 was revealed to be identical to synthetic BBS. Therefore, it is concluded from the above results that the skin of B. orientalis contains the same BBS that has been isolated from the skin of European Bombina.