Protocol for mitochondrial isolation and sub-cellular localization assay for mitochondrial proteins
Danyi Zhou, Sheng Zhong, Xinyu Han, Dandan Liu, Hezhi Fang, Ya Wang
Abstract
Here, we provide a protocol to isolate mitochondria from cultured cells and extract differently located mitochondrial proteins. We detail steps to separate both integral and peripheral membrane proteins from soluble proteins using sonication. We describe the separation of integral membrane proteins from the peripheral membrane and soluble proteins using sodium carbonate extraction. Furthermore, we detail the use of proteinase K and Triton X-100 to distinguish outer membrane proteins from mitochondrial proteins.
Topics & Concepts
Integral membrane proteinMitochondrionPeripheral membrane proteinTranslocase of the inner membraneMitochondrial membrane transport proteinMembrane proteinSonicationInner mitochondrial membraneCell biologyMitochondrial carrierChemistryMembraneTranslocase of the outer membraneBiochemistryBacterial outer membraneBiologyChromatographyGeneEscherichia coliMitochondrial Function and PathologyMetabolism and Genetic DisordersMetabolomics and Mass Spectrometry Studies