PMID- 11964259
OWN - NLM
STAT- MEDLINE
DCOM- 20020924
LR  - 20201208
IS  - 0006-3495 (Print)
IS  - 1542-0086 (Electronic)
IS  - 0006-3495 (Linking)
VI  - 82
IP  - 5
DP  - 2002 May
TI  - GTP-induced membrane binding and ion channel activity of annexin VI: is annexin 
      VI a GTP biosensor?
PG  - 2737-45
AB  - 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.
FAU - Kirilenko, Aneta
AU  - Kirilenko A
AD  - Department of Cellular Biochemistry, Nencki Institute of Experimental Biology, 
      02-093 Warsaw, Poland.
FAU - Golczak, Marcin
AU  - Golczak M
FAU - Pikula, Slawomir
AU  - Pikula S
FAU - Buchet, Rene
AU  - Buchet R
FAU - Bandorowicz-Pikula, Joanna
AU  - Bandorowicz-Pikula J
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Biophys J
JT  - Biophysical journal
JID - 0370626
RN  - 0 (Annexin A6)
RN  - 0 (Detergents)
RN  - 0 (Ion Channels)
RN  - 0 (Lipid Bilayers)
RN  - 0 (Membrane Proteins)
RN  - 0 (Recombinant Proteins)
RN  - 37589-80-3 (Guanosine 5'-O-(3-Thiotriphosphate))
RN  - 3WJQ0SDW1A (Polyethylene Glycols)
RN  - 86-01-1 (Guanosine Triphosphate)
RN  - 9002-93-1 (Octoxynol)
RN  - 9036-19-5 (Nonidet P-40)
RN  - EC 3.6.1.- (GTP-Binding Proteins)
SB  - IM
MH  - Annexin A6/isolation & purification/*metabolism
MH  - Biosensing Techniques
MH  - Circular Dichroism
MH  - Detergents
MH  - GTP-Binding Proteins/*metabolism
MH  - Guanosine 5'-O-(3-Thiotriphosphate)/*pharmacokinetics
MH  - Guanosine Triphosphate/*physiology
MH  - Humans
MH  - Ion Channels/chemistry/*physiology
MH  - Lipid Bilayers/chemistry
MH  - Membrane Proteins/*metabolism
MH  - Octoxynol
MH  - Polyethylene Glycols
MH  - Protein Conformation
MH  - Recombinant Proteins/isolation & purification/metabolism
MH  - Spectrophotometry, Infrared
PMC - PMC1302061
EDAT- 2002/04/20 10:00
MHDA- 2002/09/25 06:00
PMCR- 2003/05/01
CRDT- 2002/04/20 10:00
PHST- 2002/04/20 10:00 [pubmed]
PHST- 2002/09/25 06:00 [medline]
PHST- 2002/04/20 10:00 [entrez]
PHST- 2003/05/01 00:00 [pmc-release]
AID - S0006-3495(02)75614-2 [pii]
AID - 10.1016/S0006-3495(02)75614-2 [doi]
PST - ppublish
SO  - Biophys J. 2002 May;82(5):2737-45. doi: 10.1016/S0006-3495(02)75614-2.