Shan Zhong1, De-po Yang, Zheng Cui. 1. School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou 510080, China. lizhongshan100@yahoo.com.cn
Abstract
OBJECTIVE: To study the anticoagulant constituents in dried leech (Whitmania pigra Whitman). METHOD: The plasma recalcification time (PRT) as index, the constituents with anticoagulant activity were isolated and purified by anion-exchange chromatography on Sephadex DEAE A-50, gel permeation chromatography on Sephadex G-25 and Sephadex LH-20 columns, and then reversed phase high-performance liquid chromatography successively. RESULT: Three anticoagulant polypeptides were isolated and purified. Compounds 1 and 2 can be translated each other in natural conditions, and their molecular weights are 7100 and 5531, respectively. Compound 3 was identified as a pure polypeptide by HPLC and SDS-PAGE, and its molecular weight was determined as 8 608 by MALDI-TOF-MS. The amino acid composition of compound 3 was also determined. CONCLUSION: Compound 3 was inferred to be a novel anticoagulant, and named whitmanin.
OBJECTIVE: To study the anticoagulant constituents in dried leech (Whitmania pigra Whitman). METHOD: The plasma recalcification time (PRT) as index, the constituents with anticoagulant activity were isolated and purified by anion-exchange chromatography on Sephadex DEAE A-50, gel permeation chromatography on Sephadex G-25 and Sephadex LH-20 columns, and then reversed phase high-performance liquid chromatography successively. RESULT: Three anticoagulant polypeptides were isolated and purified. Compounds 1 and 2 can be translated each other in natural conditions, and their molecular weights are 7100 and 5531, respectively. Compound 3 was identified as a pure polypeptide by HPLC and SDS-PAGE, and its molecular weight was determined as 8 608 by MALDI-TOF-MS. The amino acid composition of compound 3 was also determined. CONCLUSION: Compound 3 was inferred to be a novel anticoagulant, and named whitmanin.