Simple purification and functional reconstitution of octopus photoreceptor Gq, which couples rhodopsin to phospholipase C

Biochemistry
S KikkawaM Tsuda

Abstract

In invertebrate photoreceptors, illuminated rhodopsin activates multiple G proteins, which are assumed to initiate multiple phototransduction cascades. In this paper, we focused on one of the phototransduction cascades, which utilizes rhodopsin, a Gq-like G protein, and phospholipase C (PLC). A Gq-like G protein from octopus photoreceptors was successfully purified to apparent homogeneity as an active form by simple two-step chromatography. The purified G protein had an alpha beta gamma-trimeric structure consisting of 44-kDa alpha, 37-kDa beta, and 9-kDa gamma subunits. The 44-kDa alpha subunit was assigned to the Gq class by western blot with antiserum against mammalian Gq alpha and by partial amino acid sequencing of its proteolytic fragments. Light-dependent binding of GTP gamma S was observed when the purified octopus Gq was reconstituted with octopus rhodopsin that had been integrated into phospholipid vesicles. Octopus Gq activated PLC beta 1 purified from bovine brain dose-dependently in the presence of A1F4-. Finally, light- and GTP-dependent activation of PLC beta 1 was observed in a reconstitution system consisting of octopus rhodopsin, Gq, and bovine PLC beta 1.

Citations

Dec 14, 1999·Molecular Neurobiology·J S LottJ B Findlay
Mar 14, 2007·Pflügers Archiv : European journal of physiology·David M Berson
Jan 11, 2000·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·C BamseyJ Mitchell
Jun 5, 1999·Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology·T SuzukiY Tsukahara
Oct 6, 2001·Progress in Retinal and Eye Research·I M Pepe
Sep 21, 2010·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Hisao Tsukamoto, Akihisa Terakita
Oct 4, 2008·Proceedings of the National Academy of Sciences of the United States of America·Mitsumasa KoyanagiAkihisa Terakita
Jun 3, 2008·Photochemistry and Photobiology·Mitsumasa Koyanagi, Akihisa Terakita
Jan 14, 2004·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Yoshinori Shichida, Takahiro Yamashita
Aug 18, 2016·PloS One·Tomohiro SugiharaAkihisa Terakita
Oct 18, 2001·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·D Arendt, J Wittbrodt

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