MRB8620 is a mitochondrial membrane-associated protein. (A) An N-terminal transmembrane domain (TMD) predicted in MRB8620. TMD predicted by TMHMM 2.0 program is located between the 90th and 119th amino acid sites with the probability of each residue being part of the TMD peaking ∼0.25. Predicted N-terminal mt localization signal (MLS) shown on the left. A stretch of amino acids with small probability of being part of a TMD extends to the 144th amino acid (dotted line) with the same score as the amino acid marked with the asterisk. Numbering of amino acid residues is from the starting methionine. (B) Western blot analysis of digitonin fractionation of cytoplasm (Cyto) and mitochondrial (Mito), as well as Triton X-100 fractionation of mt proteins into soluble and membrane fractions, containing in the supernatant (Sup) phases or pellet (Pel), respectively. Proportional volumes of each fraction were loaded as described in Material and Methods along with an equivalent amount of lysate from whole cells (Total) taken for comparison. Antibodies used are indicated on the left.
