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Molecular and Cellular Biology, April 2002, p. 2194-2203, Vol. 22, No. 7
0270-7306/02/$04.00+0     DOI: 10.1128/MCB.22.7.2194-2203.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Calcium-Dependent Assembly of Centrin-G-Protein Complex in Photoreceptor Cells

Alexander Pulvermüller,^1* Andreas Gießl,² Martin Heck,¹ Ralf Wottrich,^2, Angelika Schmitt,² Oliver Peter Ernst,¹ Hui-Woog Choe,^1,3 Klaus Peter Hofmann,¹ and Uwe Wolfrum^2*

Institut für Medizinische Physik und Biophysik, Humboldt-Universität zu Berlin, Universitätsklinikum Charité, 10098 Berlin ,¹ Institut für Zoologie, Johannes Gutenberg-Universität Mainz, 55099 Mainz, Germany ,² Department of Chemistry, Chonbuk National University, Chonju, South Korea³

Received 25 June 2001/ Returned for modification 3 August 2001/ Accepted 17 December 2001

Photoexcitation of rhodopsin activates a heterotrimeric G-protein cascade leading to cyclic GMP hydrolysis in vertebrate photoreceptors. Light-induced exchanges of the visual G-protein transducin between the outer and inner segment of rod photoreceptors occur through the narrow connecting cilium. Here we demonstrate that transducin colocalizes with the Ca²⁺-binding protein centrin 1 in a specific domain of this cilium. Coimmunoprecipitation, centrifugation, centrin overlay, size exclusion chromatography, and kinetic light-scattering experiments indicate that Ca²⁺-activated centrin 1 binds with high affinity and specificity to transducin. The assembly of centrin-G-protein complex is mediated by the ß-complex. The Ca²⁺-dependent assembly of a G protein with centrin is a novel aspect of the supply of signaling proteins in sensory cells and a potential link between molecular translocations and signal transduction in general.

Present address: ITOX, Forschungszentrum Karlsruhe, 76128 Karlsruhe, Germany.

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