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Modified electrophoretic and digestion conditions allow a simplified mass spectrometric evaluation of disulfide bonds

SYSNO ASEP	0337531
Document Type	J - Journal Article
R&D Document Type	Journal Article
Subsidiary J	Článek ve WOS
Title	Modified electrophoretic and digestion conditions allow a simplified mass spectrometric evaluation of disulfide bonds
Title	Hmotnostně spektrometrická evaluace disulfidických vazeb umožněná modifikací podmínek elektroforézy a proteinové digesce
Author(s)	Pompach, Petr (MBU-M)_{RID, ORCID} Man, Petr (MBU-M)_{RID, ORCID} Kavan, Daniel (MBU-M)_{RID, ORCID} Hofbauerová, Kateřina (MBU-M)_ORCID Kumar, Vinay (MBU-M) Bezouška, Karel (MBU-M) Havlíček, Vladimír (MBU-M)_{RID, ORCID} Novák, Petr (MBU-M)_{RID, ORCID}
Source Title	Journal of Mass Spectrometry. - : Wiley - ISSN 1076-5174 Roč. 44, č. 11 (2009), s. 1571-1578
Number of pages	8 s.
Language	eng - English
Country	GB - United Kingdom
Keywords	disulfide bond ; cystamine ; gel electrophoresis
Subject RIV	CE - Biochemistry
R&D Projects	KJB400200501 GA AV ČR - Academy of Sciences of the Czech Republic (AV ČR)
	IAA5020403 GA AV ČR - Academy of Sciences of the Czech Republic (AV ČR)
	KJB500200612 GA AV ČR - Academy of Sciences of the Czech Republic (AV ČR)
	LC545 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
	LC07017 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
CEZ	AV0Z50200510 - MBU-M (2005-2011)
UT WOS	000272150300004
DOI	10.1002/jms.1609
Annotation	Paper presents a complete sample handling protocol, which allows processing of disulfide containing proteins at basic pH. We modified the standard SDS gel electrophoresis and protein digestion conditions by the addition of an oxidative agent, cystamine. Thismodification prevented disulfide scrambling, which we otherwise observed in the samples handled according to the general protocol. Lysozyme from hen egg was used as a model protein for the development of the method. Disulfide bonds were characterized in the following proteins - human leukocyte antigen CD69, murine leukocyte receptor NKR-P1A and glycosylated β-N-acetylhexosaminidases from Aspergillus oryzae and Penicillium oxalicum
Workplace	Institute of Microbiology
Contact	Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231
Year of Publishing	2010

Number of the records: 1