Assembly of the Preprotein Receptor MOM72/MAS70 into the Protein Import Complex of the Outer Membrane of Mitochondria

Membrane integration and assembly of MOM72 from Neurospora crassa and its yeast homolog MAS70 was studied with isolated mitochondria. After synthesis in vitro, the precursors of MOM72/MAS70 are tightly folded and expose only their N-terminal amino acid residues comprising the targeting and the membrane anchor domain. Insertion of the protein into the mitochondrial outer membrane (MOM) occurs in a time- and temperature-dependent manner and is stimulated by ATP. MOM72/MAS70 is then assembled into the outer membrane MOM complex. Whereas membrane insertion occurred independently of the presence of protease-sensitive surface components, the assembly reaction depended on such components. In the MOM complex MOM72 and MAS70 were found in the neighborhood of different components in yeast and N. crassa mitochondria. MOM72 was found in association with MOM22 in N. crassa mitochondria, whereas MAS70 was in proximity to a 37-kDa component in yeast outer mitochondrial membrane. The interaction with the 37-kDa protein is important for integration of MAS70 into the MOM complex. Thus, the 37-kDa protein plays an important role in the biogenesis of MAS70.