OpenSSL SSL_connect: Connection reset by peer in connection to v2.sherpa.ac.uk:443 PuSH - Publication Server of Helmholtz Zentrum München: Improved proteome analysis of <em>Saccharomyces cerevisiae</em> mitochondria by free-flow electrophoresis.

PuSH - Publication Server of Helmholtz Zentrum München

Zischka, H. ; Weber, G.* ; Weber, P.J.* ; Posch, A.* ; Braun, R.J. ; Bühringer, D.* ; Schneider, U.* ; Nissum, M.* ; Meitinger, T. ; Ueffing, M. ; Eckerskorn, C.*

Improved proteome analysis of Saccharomyces cerevisiae mitochondria by free-flow electrophoresis.

Proteomics 3, 906-916 (2003)
Open Access Green as soon as Postprint is submitted to ZB.
The analysis of complex cellular proteomes by means of two-dimensional gel electrophoresis (2-DE) is significantly limited by the power of resolution of this technique. Although subcellular fractionation can be a fundamental first step to increase resolution, it frequently leads to preparations contaminated with other cellular structures. Here, we chose mitochondria of Saccharomyces cerevisiae to demonstrate that an integrated zone-electrophoretic purification step (ZE), with a free-flow electrophoresis device (FFE), can assist in overcoming this problem, while significantly improving their degree of purity. Whereas mitochondrial preparations isolated by means of differential centrifugation include a considerable degree of non-mitochondrial proteins (16%), this contamination could be effectually removed by the inclusion of a ZE-FFE purification step (2%). This higher degree of purity led to the identification of many more proteins from ZE-FFE purified mitochondrial protein extracts (n = 129), compared to mitochondrial protein extracts isolated by differential centrifugation (n = 80). Moreover, a marked decrease of degraded proteins was found in the ZE-FFE purified mitochondrial protein extracts. It is noteworthy that even at a low 2-DE resolution level, a four-fold higher number (17 versus 4) of presumably low abundance proteins could be identified in the ZE-FFE purified mitochondrial protein extracts. Therefore these results represent a feasible approach for an in-depth proteome analysis of mitochondria and possibly other organelles.
Publication type Article: Journal article
Document type Scientific Article
Corresponding Author
ISSN (print) / ISBN 1615-9853
e-ISSN 1615-9861
Journal Proteomics
Quellenangaben Volume: 3, Issue: 6, Pages: 906-916 Article Number: , Supplement: ,
Publisher Wiley
Non-patent literature Publications
Reviewing status Peer reviewed
Institute(s) Institute of Human Genetics (IHG)