18O-based protein quantitation: influence of separation on quantitation outcome in analysis of Staphylococcus a. enterotoxins

HAVLIŠ, Jan, Marcela ŽUPKOVÁ, Kateřina NOVÁKOVÁ and Zbyněk ZDRÁHAL. 18O-based protein quantitation: influence of separation on quantitation outcome in analysis of Staphylococcus a. enterotoxins. 2010.

Basic information

• Original name: 18O-based protein quantitation: influence of separation on quantitation outcome in analysis of Staphylococcus a. enterotoxins
• Authors: HAVLIŠ, Jan (203 Czech Republic, guarantor, belonging to the institution), Marcela ŽUPKOVÁ (203 Czech Republic, belonging to the institution), Kateřina NOVÁKOVÁ (203 Czech Republic, belonging to the institution) and Zbyněk ZDRÁHAL (203 Czech Republic, belonging to the institution).
• Edition: 2010.

Other information

• Original language: English
• Type of outcome: Conference abstract
• Field of Study: 10406 Analytical chemistry
• Country of publisher: Hungary
• Confidentiality degree: is not subject to a state or trade secret
• RIV identification code: RIV/00216224:14310/10:00049635
• Organization unit: Faculty of Science
• Keywords in English: quantitation;mass spectrometry;staphylococcal enterotoxins;separation
• Tags: AKb, mass spectrometry (MS), Quantitative Analysis, rivok

Abstract

Food contamination by enterotoxins from Staphylococcus aureus is of intense interest from side of food microbiologists and biochemists. Our work is focused on enterotoxin H (SEH). One of the main aims of our work is to reliably determine the content of SEH in whole secretome, on a background of all other staphylococcal enterotoxins that are partially sequence homologous. Mass spectrometry driven protein quantitation seems to be the most selective way. Prior to the quantitation, proteins must be separated. As the proteins are most conveniently quantified via their tryptic peptides, the separation should ensure availability of a quantifiable tryptic peptide. Here we present a study on influence of different separation ways, namely polyacrylamide gel electrophoresis, reversed phase liquid chromatography and off-gel isoelectric focusation, on availability of quantifiable tryptic peptides of SEH for 18O-based protein quantitation routine.

Links

• GA203/09/0148, research and development project: Name: Moderní metody detekce a stanovení mikrobiálních toxinů

Investor: Czech Science Foundation

• MSM0021622413, plan (intention): Name: Proteiny v metabolismu a při interakci organismů s prostředím

Investor: Ministry of Education, Youth and Sports of the CR, Proteins in metabolism and interaction of organisms with the environment

• MSM0021622415, plan (intention): Name: Molekulární podstata buněčných a tkáňových regulací

Investor: Ministry of Education, Youth and Sports of the CR, Molecular basis of cell and tissue regulations