title: Time resolution of binding and membrane insertion of a mitochondrial signal peptide: correlation with structural changes and evidence for cooperativity.
abstract: Utilizing a recently developed novel fluorescence technique [Wall et al. (1995) Mol. Membr. Biol. 12, 183-192], it is shown that the interactions of p25, the leader peptide of subunit IV of cytochrome c oxidase, with phospholipid membranes can be identified in real time. p25 is observed to bind following stopped-flow mixing of the peptide with phospholipid membranes with rate constants up to about 700 s-1 and then insert into the membrane with rate constants on the order of 0.4 s-1. Comparison of these processes with similarly time-resolved experiments performed with a stopped-flow CD spectrometer revealed that p25 does not become alpha-helical upon binding to the membrane. Following membrane insertion, however, p25 was observed to adopt an alpha-helical configuration. The temperature dependency of these processes was then found to yield activation energies for the respective components of the p25-membrane interaction.
authors.affiliation: Department of Biological & Chemical Sciences, University of Essex, Colchester, UK.
authors.forename: C
authors.initials: C
authors.lastname: Golding
authors.forename: S
authors.initials: S
authors.lastname: Senior
authors.forename: M T
authors.initials: MT
authors.lastname: Wilson
authors.forename: P
authors.initials: P
authors.lastname: O'Shea
mesh_terms: D002462:Cell Membrane / Q000378:metabolism; D002942:Circular Dichroism; D004563:Electrochemistry; D003576:Electron Transport Complex IV / Q000737:chemistry / Q000378:metabolism*; D004792:Enzyme Precursors / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D008928:Mitochondria / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D021382:Protein Sorting Signals / Q000737:chemistry / Q000378:metabolism*; D013050:Spectrometry, Fluorescence
chemical_list: D004792:Enzyme Precursors; D010743:Phospholipids; D021382:Protein Sorting Signals; D003576:Electron Transport Complex IV