Rapid single-step affinity purification of HA-tagged plant mitochondria (Plant Physiol)
In the middle part of the 20th century, cell biology leapt forward with the development of differential centrifugation methods for purifying subcellular compartments. However, these methods require large amounts of starting material and can suffer from contamination. Kuhnert et al. present a new simple and fast method for isolating plant mitochondria through affinity tagging. They generated an N-terminal translational fusion of the TOM5 gene encoding an outer-mitochondrial membrane protein with triple hemagglutin (HA) tagged synthetic GFP (sGFP). Tagged mitochondria can be purified by co-immunopurification with magnetic anti-HA beads. The authors demonstrate the usefulness of this method through a proteomic comparison of mitochondria from wild-type and the bou2 mutant that is deficient in the mitochondrial glutamate transporter À BOUT DE SOUFFLE (BOU) (“breathless”). (Summary by Mary Williams) Plant Physiol. 10.1104/pp.19.00732
