Details zur Publikation

Kategorie Textpublikation
Referenztyp Zeitschriften
DOI 10.1002/pmic.200500920
Titel (primär) Comprehensive quantitative proteome analysis of 20S proteasome subtypes from rat liver by isotope coded affinity tag and 2-D gel-based approaches
Autor Schmidt, F.; Dahlmann, B.; Janek, K.; Kloss, A.; Wacker, M.; Ackermann, R.; Thiede, B.; Jungblut, P.R.
Quelle Proteomics
Erscheinungsjahr 2006
Department PROTEOM
Band/Volume 6
Heft 16
Seite von 4622
Seite bis 4632
Sprache englisch
Keywords 20S proteasome subtypes; isotope-coded affinity tag; MS; MS-screener; quantitative proteomics
Abstract Quantitative protein profiling is an essential part of proteomics and requires technologies that accurately, reproducibly, and comprehensively identify and quantify proteins. Over the past years, many quantitative proteomic methods have been developed. Here, 20S proteasome subtypes isolated from rat were compared by four approaches based on the combination of isotopecoded affinity tag (ICAT), 2-DE, LC and ESI and MALDI MS: (i) 2-DE, (ii) ICAT/2-DE MALDI-MS, (iii) ICAT/LC-ESI-MS, (iv) ICAT/LC-MALDI-MS. A definite qualitative advantage of 2-DE gels was the separation of all known protein species, the identification of cysteine sulfoxide of alpha-4 (RC6-IS) and N-terminal acetylation of several subunits. Furthermore, quantitative differences between the standard subunits beta-2, and beta-5 and their immunosubunits were only detected by 2-DE image analysis revealing a higher replacement of standard- by immuno-beta-subunits in subtype IV It was obvious that for relative quantification only protein spot and mass peaks with a certain level of intensity displayed acceptable values of SD. However, ICAT in conjunction with LC/MALDI-MS was the most accurate method for quantification
dauerhafte UFZ-Verlinkung https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=3005
Schmidt, F., Dahlmann, B., Janek, K., Kloss, A., Wacker, M., Ackermann, R., Thiede, B., Jungblut, P.R. (2006):
Comprehensive quantitative proteome analysis of 20S proteasome subtypes from rat liver by isotope coded affinity tag and 2-D gel-based approaches
Proteomics 6 (16), 4622 - 4632 10.1002/pmic.200500920