Isolation and Quality Control of Yeast Mitochondria

Abstract

The isolation of organelles devoid of other cellular compartments is crucial for studying organellar proteomes and the localization of newly identified proteins, as well as for assessing specific organellar functions. Here, we describe a protocol for the isolation of crude and highly pure mitochondria from Saccharomyces cerevisiae and provide methods for testing the functional integrity of the isolated organelles.

Keywords: Differential centrifugation; Membrane potential; Mitochondria; Osmotic swelling; Protein import; Saccharomyces cerevisiae; Sucrose-gradient purification.