Interaction of guanosine 5'-triphosphate binding protein Gq from Sepia officinalis with illuminated rhodopsin bound to concanavalin A.
The heterotrimeric guanosine 5'-triphosphate (GTP)-binding protein Gq was suggested to couple the light receptor rhodopsin with the effector phospholipase C in visual cells of invertebrates. We indirectly linked Gq from Sepia officinalis to a concanavalin A-sepharose column via rhodopsin. Rhodopsin had been solubilized previously with 10 mM n-dodecyl-beta-maltoside from the purified photosensory membrane under illumination. All three subunits of the Gq were released almost pure by elution with 100 microM GTP. The alpha and beta subunits were identified by specific antipeptide antisera. The alpha subunit has a relative molecular mass of 46 kDa, and the beta subunit of 35 kDa. The gamma subunit corresponds to a 9 kDa polypeptide owing to the molecular mass, which is similar to the G gamma subunit of squid. The use of specific antibodies shows that neither actin nor G-protein related to transducin were in the fractions eluted with GTP or alpha-methyl mannoside. We demonstrate that all three subunits of Gq were associated with rhodopsin of invertebrates. Such use of a lectin column might be useful for further investigations of the interaction of rhodopsin and Gq.
von der Heide
W
W
Institut für Biologie II (Zoologie), Rheinisch-Westfälische Technische Hochschule Aachen, Germany.
Casaretto
M
M
Rack
M
M
Stieve
H
H
D000595:Amino Acid Sequence; D000818:Animals; D002846:Chromatography, Affinity; D003208:Concanavalin A / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008362:Mannosides / Q000378:metabolism; D008766:Methylmannosides; D008969:Molecular Sequence Data; D008974:Mollusca / Q000378:metabolism*; D012243:Rhodopsin / Q000378:metabolism*
D008362:Mannosides; D008766:Methylmannosides; D003208:Concanavalin A; C008466:methylmannoside; D012243:Rhodopsin; D019204:GTP-Binding Proteins