Self-association and precursor protein binding of Saccharomyces cerevisiae Tom40p, the core component of the protein translocation channel of the mitochondrial outer membrane

The Biochemical Journal
D M GordonD Pain

Abstract

The precursor protein translocase of the mitochondrial outer membrane (Tom) is a multi-subunit complex containing receptors and a general import channel, of which the core component is Tom40p. Nuclear-encoded mitochondrial precursor proteins are first recognized by surface receptors and then pass through the import channel. The Tom complex has been purified; however, the protein-protein interactions that drive its assembly and maintain its stability have been difficult to study. Here we show that Saccharomyces cerevisiae Tom40p expressed in bacteria and purified to homogeneity associates efficiently with itself. The self-association is very strong and can withstand up to 4 M urea or 1 M salt. The tight self-association does not require the N-terminal segment of Tom40p. Furthermore, purified Tom40p preferentially recognizes the targeting sequence of mitochondrial precursor proteins. Although the binding of the targeting sequence to Tom40p is inhibited by urea concentrations in excess of 1 M, it is moderately resistant to 1 M salt. Simultaneous self-assembly and precursor protein binding suggest that Tom40p contains at least two different domains mediating these processes. The experimental approach described here should be useful...Continue Reading

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Citations

Nov 27, 2009·Journal of Molecular Medicine : Official Organ of the Gesellschaft Deutscher Naturforscher Und Ärzte·Wonchung LimHyeseong Cho
Jun 14, 2005·Mechanisms of Ageing and Development·Bartosz Szczesny, Sankar Mitra
Oct 18, 2002·Current Opinion in Cell Biology·Nikolaus Pfanner, Nils Wiedemann
Sep 13, 2011·The Journal of Biological Chemistry·Alok PandeyDebkumar Pain
Jul 19, 2008·The Journal of Biological Chemistry·Bartosz SzczesnySankar Mitra
Apr 18, 2006·Nucleic Acids Research·Ranajoy ChattopadhyaySankar Mitra
Sep 10, 2003·Proceedings of the National Academy of Sciences of the United States of America·Bartosz SzczesnyIstvan Boldogh
Jul 27, 2010·European Journal of Cell Biology·Xin Cheng, Andreas S Ivessa
Oct 26, 2002·The Journal of Biological Chemistry·Rebecca D TaylorFrank E Nargang
Aug 23, 2002·Biochimica Et Biophysica Acta·Nikolaus Pfanner, Agnieszka Chacinska

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