Interaction of presequence peptides with human translocase of inner membrane of mitochondria Tim23

Biochem Biophys Res Commun. 2013 Jul 26;437(2):292-9. doi: 10.1016/j.bbrc.2013.06.070. Epub 2013 Jun 28.

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. Tim23p, the core component of TIM23 complex, forms the import pore across the inner membrane. However, the interaction between presequence peptides and Tim23p remains unclear. Herein, we investigated the interaction of presequence peptides with the intermembrane space domain of Tim23p (Tim23IMS) by fluorescence and micro-Raman spectroscopy. The fluorescence quenching revealed that the interaction between Tim23IMS and presequence peptides is mainly electrostatic interaction. Micro-Raman spectroscopy and ANS binding experiments showed that presequence peptides induce a more compact conformation of Tim23IMS. GST pull-down experiments and tryptophan fluorescence indicated that there is no interaction between Tim23IMS and Tim50IMS.

Keywords: Interaction; Mitochondria; Presequence peptides; TIM23 complex; Tim23p.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Humans
  • Mitochondrial Membrane Transport Proteins / metabolism*
  • Mitochondrial Membranes / enzymology*
  • Mitochondrial Precursor Protein Import Complex Proteins
  • Molecular Sequence Data
  • Peptidyl Transferases / metabolism*
  • Spectrometry, Fluorescence
  • Spectrum Analysis, Raman

Substances

  • Mitochondrial Membrane Transport Proteins
  • Mitochondrial Precursor Protein Import Complex Proteins
  • TIMM23 protein, human
  • Peptidyl Transferases