J Vet Sci.  2013 Jun;14(2):107-114. 10.4142/jvs.2013.14.2.107.

Sporozoite proteome analysis of Cryptosporidium parvum by one-dimensional SDS-PAGE and liquid chromatography tandem mass spectrometry

• Siddiki AM

• KMID: 1705511
• DOI: http://doi.org/10.4142/jvs.2013.14.2.107

Abstract

Despite the development of new technologies, new challenges still remain for large scale proteomic profiling when dealing with complex biological mixtures. Fractionation prior to liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis is usually the preferred method to reduce the complexity of any biological sample. In this study, a gel LC-MS/MS approach was used to explore the stage specific proteome of Cryptosporidium (C.) parvum. To accomplish this, the sporozoite protein of C. parvum was first fractionated using SDS-PAGE with subsequent LC-MS/MS analysis. A total of 135 protein hits were recorded from 20 gel slices (from same gel lane), with many hits occurring in more than one band. Excluding all non-Cryptosporidium entries and proteins with multiple hits, 33 separate C. parvum entries were identified during the study. The overall goal of this study was to reduce sample complexity by protein fractionation and increase the possibility of detecting proteins present in lower abundance in a complex protein mixture.

Keyword

bioinformatics; Cryptosporidium; proteomics; SDS-PAGE; tandem mass spectrometry

MeSH Terms

Chemical Fractionation/methods
Chromatography, Liquid/methods/veterinary
Cryptosporidium parvum/*chemistry/growth & development/metabolism
Electrophoresis, Polyacrylamide Gel/methods/veterinary
Gene Expression Profiling/*methods/veterinary
Proteome/analysis
Proteomics/*methods
Protozoan Proteins/*analysis
Sporozoites/chemistry/metabolism
Tandem Mass Spectrometry/methods/veterinary
Proteome
Protozoan Proteins