PMID- 19153686
OWN - NLM
STAT- MEDLINE
DCOM- 20090319
LR  - 20131121
IS  - 1064-3745 (Print)
IS  - 1064-3745 (Linking)
VI  - 528
DP  - 2009
TI  - 16-BAC/SDS-PAGE analysis of membrane proteins of yeast mitochondria purified by 
      free flow electrophoresis.
PG  - 83-107
LID - 10.1007/978-1-60327-310-7_6 [doi]
AB  - Mitochondria are essential organelles in cellular metabolism. These organelles 
      are bounded by two membranes, the outer and inner membrane. Especially the inner 
      membrane comprises a high content of proteins, for example, the protein complexes 
      of the respiratory chain. High-resolution separation and analysis of such 
      membrane proteins, for example, by two-dimensional gel electrophoresis (2-DE), is 
      hampered by their hydrophobicity and tendency for aggregation. Here, we describe 
      the separation of mitochondrial membrane proteins of Saccharomyces cerevisiae by 
      16-benzyldimethyl-n-hexadecylammonium chloride/sodium dodecyl sulfate 
      polyacrylamide gel electrophoresis (16-BAC/SDS-PAGE). This method enables the 
      separation of membrane proteins owing to the solubilizing power of the ionic 
      detergents 16-BAC and SDS, respectively. Mitochondria were isolated from yeast 
      cultures by differential centrifugation and were further purified by free flow 
      electrophoresis (FFE) in zone-electrophoretic mode (ZE). Subsequently, membrane 
      proteins from ZE-FFE-purified mitochondria were enriched by carbonate extraction 
      and subjected to 16-BAC/SDS-PAGE. The resulting protein spot patterns were 
      visualized by a highly sensitive fluorescence stain with 
      ruthenium-II-bathophenantroline disulfonate chelate (RuBP), and by colloidal 
      Coomassie staining. Proteins were identified by Maldi-Tof mass spectrometry and 
      peptide mass fingerprinting.
FAU - Braun, Ralf J
AU  - Braun RJ
AD  - GSF-National Research Centre for Environment and Health, Institute of Human 
      Genetics, Munich, Germany.
FAU - Kinkl, Norbert
AU  - Kinkl N
FAU - Zischka, Hans
AU  - Zischka H
FAU - Ueffing, Marius
AU  - Ueffing M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (Carbonates)
RN  - 0 (Fatty Alcohols)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Membrane Proteins)
RN  - 0 (Quaternary Ammonium Compounds)
RN  - 0 (Saccharomyces cerevisiae Proteins)
RN  - 368GB5141J (Sodium Dodecyl Sulfate)
RN  - 45P3261C7T (sodium carbonate)
RN  - 7UI0TKC3U5 (Ruthenium)
RN  - 85474O1N9D (cetalkonium chloride)
SB  - IM
MH  - Carbonates/chemistry
MH  - Centrifugation
MH  - Chemical Fractionation/*methods
MH  - Electrophoresis, Capillary
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Fatty Alcohols
MH  - Fluorescent Dyes
MH  - Membrane Proteins/*analysis
MH  - Mitochondria/*chemistry
MH  - Quaternary Ammonium Compounds/chemistry
MH  - Ruthenium/chemistry
MH  - Saccharomyces cerevisiae/*chemistry
MH  - Saccharomyces cerevisiae Proteins/*analysis
MH  - Sodium Dodecyl Sulfate/chemistry
EDAT- 2009/01/21 09:00
MHDA- 2009/03/20 09:00
CRDT- 2009/01/21 09:00
PHST- 2009/01/21 09:00 [entrez]
PHST- 2009/01/21 09:00 [pubmed]
PHST- 2009/03/20 09:00 [medline]
AID - 10.1007/978-1-60327-310-7_6 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2009;528:83-107. doi: 10.1007/978-1-60327-310-7_6.