Molecular characterization of the alpha-subunit of Trichomonas vaginalis hydrogenosomal succinyl CoA synthetase.
The anaerobic, parasitic protist, Trichomonas vaginalis, is characterized by the absence of mitochondria and the presence of double membrane bound organelles called hydrogenosomes. Succinyl-coenzyme A synthetase is a hydrogenosomal enzyme which catalyzes the formation of ATP via substrate-level phosphorylation. We have characterized genes encoding the alpha subunit of the hydrogenosomal protein succinyl-coenzyme A synthetase (SCS). The alpha-SCS of T. vaginalis is encoded by a multigene family composed of 3 similar genes that do not appear allelic. These 3 alpha-SCS genes encode a protein with a calculated molecular mass of approximately 32.5 kDa that has > 50% identity (> 70% similarity with alpha-SCSs from Escherichia coli, Thermus flavus, and rat liver mitochondria. Antibodies raised against recombinant T vaginalis alpha-SCS expressed in bacteria were used to isolate alpha-SCS proteins from purified hydrogenosomes. These proteins partition into the soluble fraction of hydrogenosomes treated with sodium carbonate at high pH, consistent with a matrix localization in the organelle. Amino-terminal sequencing of purified alpha-SCS proteins shows that mature proteins lack a short, leader sequence of 9 amino acids. These amino terminal sequences which are cleaved from T. vaginalis alpha-SCSs are similar to each other and to all other leader sequences identifed on hydrogenosomal proteins.
Lahti
C J
CJ
Department of Microbiology and Immunology, University of California, Los Angeles, 90024-1747.
Bradley
P J
PJ
Johnson
P J
PJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D016054:DNA, Protozoan / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D017125:Genes, Protozoan; D008969:Molecular Sequence Data; D015388:Organelles / Q000201:enzymology; D011487:Protein Conformation; D051381:Rats; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013389:Succinate-CoA Ligases / Q000737:chemistry / Q000235:genetics*; D013824:Thermus / Q000201:enzymology / Q000235:genetics; D014246:Trichomonas vaginalis / Q000201:enzymology* / Q000235:genetics*
D017931:DNA Primers; D016054:DNA, Protozoan; D013389:Succinate-CoA Ligases