ATP binding site on the C-terminus of the vanilloid receptor.
Transient receptor potential channel vanilloid receptor subunit 1 (TRPV1) is a thermosensitive cation channel activated by noxious heat as well as a wide range of chemical stimuli. Although ATP by itself does not directly activate TRPV1, it was shown that intracellular ATP increases its activity by directly interacting with the Walker A motif residing on the C-terminus of TRPV1. In order to identify the amino acid residues that are essential for the binding of ATP to the TRPV1 channel, we performed the following point mutations of the Walker A motif: P732A, D733A, G734A, K735A, D736A, and D737A. Employing bulk fluorescence measurements, namely a TNP-ATP competition assay and FITC labelling and quenching experiments, we identified the key role of the K735 residue in the binding of the nucleotide. Experimental data was interpreted according to our molecular modelling simulations.
Grycova
Lenka
L
Institute of Physiology, Academy of Sciences of the Czech Republic, Vídenská 1083, 14220 Prague, Czech Republic.
Lansky
Zdenek
Z
Friedlova
Eliska
E
Vlachova
Viktorie
V
Kubala
Martin
M
Obsilova
Veronika
V
Obsil
Tomas
T
Teisinger
Jan
J
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000737:chemistry*; D001665:Binding Sites; D003198:Computer Simulation; D008956:Models, Chemical; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation; D050916:TRPV Cation Channels / Q000737:chemistry* / Q000648:ultrastructure*
D050916:TRPV Cation Channels; C494484:TRPV1 receptor; C045674:2',3'-O-(2,4,6-trinitro-cyclohexadienylidine)adenosine 5'-triphosphate; D000255:Adenosine Triphosphate