[Studies on anticoagulant constituents in dried Whitmania pigra].
OBJECTIVE
To study the anticoagulant constituents in dried leech (Whitmania pigra Whitman).


METHODS
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.


RESULTS
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.


CONCLUSIONS
Compound 3 was inferred to be a novel anticoagulant, and named whitmanin.
Zhong
Shan
S
School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou 510080, China. lizhongshan100@yahoo.com.cn
Yang
De-po
DP
Cui
Zheng
Z
D000818:Animals; D000925:Anticoagulants / Q000032:analysis* / Q000737:chemistry* / Q000302:isolation & purification; D007865:Leeches / Q000737:chemistry*; D008970:Molecular Weight
D000925:Anticoagulants