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Originally published In Press as doi:10.1074/mcp.M700321-MCP200 on December 18, 2007.
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Molecular & Cellular Proteomics 7:785-799, 2008.
© 2008 by The American Society for Biochemistry and Molecular Biology, Inc.

Research

Quantitative Analysis of Snake Venoms Using Soluble Polymer-based Isotope Labeling*,S

Jacob A. Galan{ddagger}, Minjie Guo{ddagger}, Elda E. Sanchez§, Esteban Cantu§, Alexis Rodriguez-Acosta, John C. Perez§ and W. Andy Tao{ddagger},||

From the {ddagger} Departments of Biochemistry, Chemistry, Medicinal Chemistry, and Molecular Pharmacology, Purdue University, West Lafayette, Indiana 47907, § Natural Toxins Research Center, College of Arts and Science, Texas A&M University-Kingsville, Kingsville, Texas 78363, and Immunochemistry Section, Tropical Medicine Institute, Universidad Central de Venezuela, Caracas 1041, Venezuela

We present the design and synthesis of a new quantitative strategy termed soluble polymer-based isotope labeling (SoPIL) and its application as a novel and inclusive method for the identification and relative quantification of individual proteins in complex snake venoms. The SoPIL reagent selectively captures and isolates cysteine-containing peptides, and the subsequent tagged peptides are released and analyzed using nanoflow liquid chromatography-tandem mass spectrometry. The SoPIL strategy was used to quantify venom proteins from two pairs of venomous snakes: Crotalus scutulatus scutulatus type A, C. scutulatus scutulatus type B, Crotalus oreganus helleri, and Bothrops colombiensis. The hemorrhagic, hemolytic, clotting ability, and fibrinogenolytic activities of crude venoms were measured and correlated with difference in protein abundance determined by the SoPIL analysis. The SoPIL approach could provide an efficient and widely applicable tool for quantitative proteomics.

|| To whom correspondence should be addressed. Tel.: 765-494-9605; E-mail: watao{at}purdue.edu


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