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Research articles

ScienceAsia (): 273-277 |doi: 10.2306/scienceasia1513-1874...273


pH gradient electrophoresis and biological activity analysis of proteins from Malayan pit viper (Calloselasma rhodostoma) venom


Piboon Pornmaneea,*, John C. P˙rezb, Elda E. S˙nchezb, Orawan Khowc, Narumol Pakmaneec, Pannipa Chulasugandhac, Lawan Chanhomec, Amorn Petsomd

 
ABSTRACT:     The Malayan pit viper (Calloselasma rhodostoma) is a snake found in most of Southeast Asia. The snake˙s venom contains proteins with various biological effects. In this study, proteins from Malayan pit viper venom were analysed by electrophoresis titration (ET) and two dimensional gel electrophoresis (2-D gel). In addition, venom proteins were separated by high performance liquid chromatography (HPLC) connected to a hydrophobic interactive chromatography (HIC) column. Fractions collected from HPLC were tested for biological activities. As the result, the ET profile showed that crude venom consisted of both positively and negatively charged proteins. Most of the 191 protein spots found on 2-D gel of crude venom have an isoelectric point in the range 4.5˙5.5. After HPLC, eighteen fractions were eluted from HIC column. Each fraction was tested for fibrinolytic, haemorrhagic, gelatinase, and disintegrin activities. Both fibrinolytic and haemorrhagic fractions showed gelatinase activity as well, while the fibrinolytic fraction had no haemorrhagic activity. Our results are valuable to venom research and drug discovery.

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a Program of Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand
b Natural Toxins Research Center, Texas A&M University-Kingsville, MSC 158, Kingsville, Texas, 78363, USA
c Queen Saovabha Memorial Institute, Thai Red Cross Society, Bangkok 10330, Thailand
d Institute of Biotechnology and Genetic Engineering, Chulalongkorn University, Bangkok 10330, Thailand

* Corresponding author, E-mail: boon9165@hotmail.com

Received 21 Jan 2008, Accepted 24 Jun 2008