Display Settings:

Format

Send to:

Choose Destination
See comment in PubMed Commons below
Biophys J. 2002 May;82(5):2737-45.

GTP-induced membrane binding and ion channel activity of annexin VI: is annexin VI a GTP biosensor?

Author information

  • 1Department of Cellular Biochemistry, Nencki Institute of Experimental Biology, 02-093 Warsaw, Poland.

Abstract

Annexin VI (AnxVI) formed ion channels in planar lipid bilayers that were induced by the addition of millimolar guanosine 5'-triphosphate (GTP) at pH 7.4 and that were not accompanied by a penetration of the protein into the membrane hydrophobic region. GTP-influenced interactions of AnxVI with Ca2+/liposomes produced small structural alterations as revealed by circular dichroism and infrared spectroscopies. Guanosine 5'-3-O-(thio)-triphosphate (GTPgammaS) binding to AnxVI, promoted by the photorelease of GTPgammaS from GTPgammaS[1-(4,5-dimethoxy-2-nitrophenyl)-ethyl] (caged-GTPgammaS), affected three to four amino acid residues of AnxVI in the presence of Ca2+/liposomes, while about eight or nine amino acid residues were altered in their absence. This suggested that the nucleotide-binding site overlapped the lipid-binding domain of AnxVI. The binding of the fluorescent GTP analog, 2'-(or 3')-O-(2,4,6-trinitrophenyl)guanosine 5'-triphosphate (TNP-GTP) to AnxVI was optimal in the presence of Ca2+/liposomes, with a dissociation constant (K(d)) of 1 microM and stoichiometry of 1. TNP-GTP promoted fluorescence resonance energy transfer from tryptophan residues to the nucleotide. Ion conductance and fluorescence measurements of the C- and N-terminal fragments of AnxVI indicated distinct GTP-binding properties, suggesting that the existence of the GTP-induced ion channel activity of AnxVI is associated with the flexibility of the two halves of the protein. Such structural flexibility could contribute to a molecular mechanism of AnxVI acting as a GTP biosensor.

PMID:
11964259
[PubMed - indexed for MEDLINE]
PMCID:
PMC1302061
Free PMC Article
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Elsevier Science Icon for PubMed Central
    Loading ...
    Write to the Help Desk