title: Interaction of divalent metal ions with human translocase of inner membrane of mitochondria Tim50.
abstract: The preprotein translocase of the inner membrane of mitochondria (TIM23 complex) is the main entry gate for proteins of the matrix and the inner membrane. Tim50 is a major receptor for transporting the precursor protein across the mitochondrial inner membrane in TIM23 complex. However, the interaction of divalent metal ions with Tim50 and the contribution in the interaction of presequence peptide with Tim50 are still unknown. Herein, we investigated the interaction of divalent metal ions with the intermembrane space domain of Tim50 (Tim50(IMS)) and the interaction of presequence peptides with Tim50(IMS) in presence of Ca(2+) ion by fluorescence spectroscopy in vitro. The static fluorescence quenching indicates the existence of strong binding between divalent metal ions and Tim50(IMS). The order of the binding strength is Ca(2+), Mg(2+), Cu(2+), Mn(2+), and Co(2+) (from strong to weak). Moreover, the interaction of presequence peptides with Tim50(IMS) is weakened in presence of Ca(2+) ion, which implicates that Ca(2+) ion may play an important role in the protein import by TIM23 complex.
authors.affiliation: School of Physics Science, Department of Biophysics, Nankai University, Tianjin 300071, PR China.
authors.forename: Yongqiang
authors.initials: Y
authors.lastname: Zhang
authors.forename: Honghua
authors.initials: H
authors.lastname: Deng
authors.forename: Xinzheng
authors.initials: X
authors.lastname: Zhang
authors.forename: Shu Jie
authors.initials: SJ
authors.lastname: Li
mesh_terms: D002118:Calcium / Q000737:chemistry*; D002413:Cations, Divalent / Q000737:chemistry; D006801:Humans; D026901:Membrane Transport Proteins / Q000737:chemistry*; D033681:Mitochondrial Membrane Transport Proteins / Q000737:chemistry; D000090622:Mitochondrial Precursor Protein Import Complex Proteins; D017434:Protein Structure, Tertiary
chemical_list: D002413:Cations, Divalent; D026901:Membrane Transport Proteins; D033681:Mitochondrial Membrane Transport Proteins; D000090622:Mitochondrial Precursor Protein Import Complex Proteins; C438387:TIMM23 protein, human; C509348:TIMM50 protein, human; D002118:Calcium