Trichomonas vaginalis Hmp35, a putative pore-forming hydrogenosomal membrane protein, can form a complex in yeast mitochondria.
An abundant integral membrane protein, Hmp35, has been isolated from hydrogenosomes of Trichomonas vaginalis. This protein has no known homologue and exists as a stable 300-kDa complex, termed HMP35, in membranes of the hydrogenosome. By using blue native gel electrophoresis, we found the HMP35 complex to be stable in 2 m NaCl and up to 5 m urea. The endogenous Hmp35 protein was largely protease-resistant. The protein has a predominantly beta-sheet structure and predicted transmembrane domains that may form a pore. Interestingly, the protein has a high number of cysteine residues, some of which are arranged in motifs that resemble the RING finger, suggesting that they could be coordinating zinc or another divalent cation. Our data show that Hmp35 forms one intramolecular but no intermolecular disulfide bonds. We have isolated the HMP35 complex by expressing a His-tagged Hmp35 protein in vivo followed by purification with nickel-agarose beads. The purified 300-kDa complex consists of mostly Hmp35 with lesser amounts of 12-, 25-27-, and 32-kDa proteins. The stoichiometry of proteins in the complex indicates that Hmp35 exists as an oligomer. Hmp35 can be targeted heterologously into yeast mitochondria, despite the lack of homology with any yeast protein, demonstrating the compatibility of mitochondrial and hydrogenosomal protein translocation machineries.
Dyall
Sabrina D
SD
Department of Microbiology, Immunology, and Molecular Genetics, UCLA, Los Angeles, California 90095-1489 and the Department of Chemistry and Biochemistry and the Molecular Biology Institute, UCLA, Los Angeles, California 90095-1569.
Lester
Dianna C
DC
Schneider
Rachel E
RE
Delgadillo-Correa
Maria G
MG
Plümper
Evelyn
E
Martinez
Ariadna
A
Koehler
Carla M
CM
Johnson
Patricia J
PJ
D000595:Amino Acid Sequence; D000818:Animals; D003545:Cysteine / Q000235:genetics; D004220:Disulfides / Q000378:metabolism; D010450:Endopeptidases / Q000494:pharmacology; D006859:Hydrogen / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D017433:Protein Structure, Secondary; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D014246:Trichomonas vaginalis / Q000235:genetics* / Q000378:metabolism*
D004220:Disulfides; D008565:Membrane Proteins; D015800:Protozoan Proteins; D006859:Hydrogen; D010450:Endopeptidases; D003545:Cysteine