title: Partially folded rhodanese or its N-terminal sequence can disrupt phospholipid vesicles.
abstract: Rhodanese (thiosulfate cyanide sulfurtransferase; E.C. 2.8.1.1) is a mitochondrial enzyme that is unprocessed after import. We describe in vitro experiments showing that partially folded rhodanese can interact with lipid bilayers. The interaction was monitored by measuring the ability of rhodanese to disrupt small unilamellar vesicles composed of phosphatidylserine and to release 6-carboxyfluorescein that was trapped in the liposomes. Partially folded rhodanese, derived by dilution of urea-unfolded enzyme, efficiently induced liposome leakage. Native rhodanese had no effect on liposome integrity. Liposome disruption progressively decreased as rhodanese was given the opportunity to refold or aggregate before introduction of the liposomes. A synthetic 23 amino acid peptide representing the N-terminal sequence of rhodanese was very efficient at disrupting the liposomes. Shorter peptides chosen from within this sequence (residues 11-23 or residues 1-17) had no effect on liposome disruption. A peptide representing the tether region that connects the domains of the enzyme was also without effect. These results are consistent with the hypothesis that the N-terminal sequence of rhodanese is an uncleaved leader sequence, and can interact with membrane components that are involved in the mitochondrial uptake of this protein.
authors.affiliation: Department of Biochemistry, University of Texas Health Science Center, San Antonio 78284-7760.
authors.forename: J A
authors.initials: JA
authors.lastname: Mendoza
authors.forename: E
authors.initials: E
authors.lastname: Grant
authors.forename: P M
authors.initials: PM
authors.lastname: Horowitz
mesh_terms: D000595:Amino Acid Sequence; D008051:Lipid Bilayers; D008081:Liposomes; D008969:Molecular Sequence Data; D010743:Phospholipids / Q000737:chemistry*; D017510:Protein Folding; D013884:Thiosulfate Sulfurtransferase / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D008051:Lipid Bilayers; D008081:Liposomes; D010743:Phospholipids; D013884:Thiosulfate Sulfurtransferase