Trapping oxidative folding intermediates during translocation to the intermembrane space of mitochondria: in vivo and in vitro studies.
The MIA40 pathway is a novel import pathway in mitochondria specific for cysteine-rich proteins of the intermembrane space (IMS). The newly synthesised precursors are trapped in the IMS by a disulfide relay mechanism that involves introduction of disulfides from the sulfhydryl oxidase Erv1 to the redox-regulated import receptor Mia40 and then on to the substrate. This thiol-disulfide exchange mechanism is essential for the import and oxidative folding of the incoming cysteine-rich substrate proteins. In this chapter we will describe the experimental methods that have been developed in order to study and characterise disulfide-trapped intermediates in yeast mitochondria.
Sideris
Dionisia P
DP
Department of Biology, University of Crete and Institute of Molecular Biology and Biotechnology-Foundation for Research and Technology Hellas, Heraklion, Crete, Greece.
Tokatlidis
Kostas
K
D001345:Autoradiography; D004586:Electrophoresis; D004591:Electrophoresis, Polyacrylamide Gel; D007425:Intracellular Membranes / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism*; D008954:Models, Biological; D010084:Oxidation-Reduction; D017510:Protein Folding; D021381:Protein Transport / Q000502:physiology; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D015003:Yeasts / Q000378:metabolism
D011506:Proteins