Synthesis and sorting of mitochondrial translation products.
Mitochondria are essential organelles of eukaryotic cells. The biogenesis of mitochondria depends on the coordinated function of two separate genetic systems: one in the nucleus and one in the organelle. The study of mitochondria requires the analysis of both genetic systems and their protein products. In this chapter, we focus on the translation and sorting of mitochondrially encoded proteins into the mitochondrial inner membrane in the baker's yeast Saccharomyces cerevisiae. The starting point is the labeling of these proteins, followed by some of the methods developed to investigate their topology and membrane incorporation. The methods described here can be applied also to the study of other aspects of organelle biogenesis such as folding, assembly, and degradation of proteins.
Bauerschmitt
Heike
H
Adolf-Butenandt-Institute of Physiological Chemistry, University of Munich, Munich, Germany.
Funes
Soledad
S
Herrmann
Johannes M
JM
D002254:Carbonates; D002498:Centrifugation; D004591:Electrophoresis, Polyacrylamide Gel; D019286:Endopeptidase K / Q000378:metabolism; D008565:Membrane Proteins / Q000302:isolation & purification; D008928:Mitochondria / Q000378:metabolism*; D051336:Mitochondrial Membranes / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000302:isolation & purification / Q000378:metabolism*; D008967:Molecular Biology / Q000379:methods*; D009995:Osmosis; D014176:Protein Biosynthesis; D021381:Protein Transport; D012394:Rosaniline Dyes; D012441:Saccharomyces cerevisiae / Q000254:growth & development / Q000378:metabolism*; D013194:Staining and Labeling
D002254:Carbonates; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D012394:Rosaniline Dyes; C048139:Coomassie blue; D019286:Endopeptidase K