Identification of a tubulin binding motif on the P2X2 receptor.
To isolate proteins interacting with P2X receptors, GST fusion proteins containing the intracellular C terminal tail of P2X(2), P2X(5), or P2X(7) were used as bait to screen detergent extracts of rat brain synaptosomes. By SDS-PAGE combined with mass spectrometry, two interacting proteins were identified: betaIII tubulin and myelin basic protein. While myelin basic protein bound to all three P2X subunits, betaIII tubulin interacted exclusively with the P2X(2) subunit. The tubulin binding domain could be confined to a proline-rich segment (amino acids 371-412) of the P2X(2) subunit. Our results suggest a role for microtubules in the cellular localisation of the P2X(2) receptor.
Gendreau
Sandra
S
Department of Molecular Pharmacology, Medical School of the Technical University of Aachen, Wendlingweg 2, D-52074, Aachen, Germany.
Schirmer
Jörg
J
Schmalzing
Günther
G
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D018076:DNA, Complementary; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D011485:Protein Binding; D051381:Rats; D017207:Rats, Sprague-Dawley; D018048:Receptors, Purinergic P2 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D058477:Receptors, Purinergic P2X3; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013574:Synaptosomes / Q000378:metabolism; D014404:Tubulin / Q000378:metabolism*
D018076:DNA, Complementary; D003902:Detergents; C546410:P2rx3 protein, rat; D018048:Receptors, Purinergic P2; D058477:Receptors, Purinergic P2X3; D011993:Recombinant Fusion Proteins; D014404:Tubulin