Quantitative analysis of snake venoms using soluble polymer-based isotope labeling.
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.
Galan
Jacob A
JA
Department of Biochemistry, Purdue University, West Lafayette, Indiana 47907, USA.
Guo
Minjie
M
Sanchez
Elda E
EE
Cantu
Esteban
E
Rodriguez-Acosta
Alexis
A
Perez
John C
JC
Tao
W Andy
WA
D000818:Animals; D017837:Bothrops; D017839:Crotalus; D006801:Humans; D007553:Isotope Labeling / Q000379:methods*; D049329:Nanostructures; D010455:Peptides / Q000032:analysis* / Q000302:isolation & purification; D011108:Polymers / Q000737:chemistry; D040901:Proteomics; D012910:Snake Venoms / Q000737:chemistry*; D012995:Solubility
D010455:Peptides; D011108:Polymers; D012910:Snake Venoms