Interaction of the precursor to mitochondrial aspartate aminotransferase and its presequence peptide with model membranes

Donate, F; Yanez, AJ; Iriarte, A; Martinez-Carrion, M

Abstract

The possible contribution of the mature portion of a mitochondrial precursor protein to its interaction with membrane lipids is unclear. To address this issue, we examined the interaction of the precursor to mitochondrial aspartate aminotransferase (pmAAT) and of a synthetic peptide corresponding to the 29-residue presequence peptide (mAAT-pp) with anionic phospholipid vesicles. The affinity of mAAT-pp and pmAAT for anionic vesicles is nearly identical. Results obtained by analyzing the effect of mAAT-pp or full-length pmAAT on either the permeability or microviscosity of the phospholipid vesicles are consistent with only a shallow insertion of the presequence peptide in the bilayer. Analysis of the quenching of Trp-17 fluorescence by brominated phospholipids reveals that this presequence residue inserts to a depth of approximately 9 Angstrom hom the center of the bilayer, Furthermore, in membrane-bound pmAAT or mAAT-pp, both Arg-8 and Arg-28 are accessible to the solvent. These results suggest that the presequence segment lies close to the surface of the membrane and that the mature portion of the precursor protein has little effect on the affinity or mode of binding of the presequence to model membranes. Tn the presence of vesicles, mAAT-pp adopts considerable a-helical structure. Hydrolysis by trypsin after Arg-g results in the dissociation of the remaining 21-residue C-terminal peptide fragment from the membrane bilayer, suggesting that the N-terminal portion of the presequence is essential for membrane binding. Based on these results, we propose that the presequence peptide may contain dual recognition elements for both the lipid and import receptor components of the mitochondrial membrane.

Más información

Título según WOS: Interaction of the precursor to mitochondrial aspartate aminotransferase and its presequence peptide with model membranes
Título según SCOPUS: Interaction of the precursor to mitochondrial aspartate aminotransferase and its presequence peptide with model membranes
Título de la Revista: JOURNAL OF BIOLOGICAL CHEMISTRY
Volumen: 275
Número: 44
Editorial: AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
Fecha de publicación: 2000
Página de inicio: 34147
Página final: 34156
Idioma: English
URL: http://www.jbc.org/cgi/doi/10.1074/jbc.M004494200
DOI:

10.1074/jbc.M004494200

Notas: ISI, SCOPUS