Determination of yeast mitochondrial KHE activity, osmotic swelling and mitophagy.
The mitochondrial K(+)/H(+) exchanger (KHE) is a key regulator of mitochondrial K(+), the most abundant cellular cation, and thus for volume control of the organelle. Downregulation of the mitochondrial KHE results in osmotic swelling and autophagic degradation of the organelle. This chapter describes methods to shut-off expression of Mdm38p, an essential factor of the mitochondrial KHE, and to observe the cellular consequences thereof, in particular changes in KHE activity and morphogenetic changes of mitochondria by applying new techniques developed in our laboratories.
Max F. Perutz Laboratories, Department of Genetics, University of Vienna, Campus Vienna Biocenter, Wien, Austria.
Karin
K
Nowikovsky
Rodney J
RJ
Devenish
Elisabeth
E
Froschauer
Rudolf J
RJ
Schweyen
D001343:Autophagy; D015374:Biosensing Techniques / Q000379:methods*; D005786:Gene Expression Regulation; D053078:Membrane Potential, Mitochondrial; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000378:metabolism; D033681:Mitochondrial Membrane Transport Proteins / Q000378:metabolism*; D024101:Mitochondrial Proteins; D009995:Osmosis; D011188:Potassium / Q000378:metabolism; D027961:Potassium-Hydrogen Antiporters / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*
C488529:MDM38 protein, S cerevisiae; D008565:Membrane Proteins; D033681:Mitochondrial Membrane Transport Proteins; D024101:Mitochondrial Proteins; D027961:Potassium-Hydrogen Antiporters; D029701:Saccharomyces cerevisiae Proteins; D011188:Potassium