Bovine Retinal Nucleoside Diphosphate Kinase: Biochemistry and Molecular Cloning

Published

August 1, 2000

Author(s)

N G. Abdulaev, D Kakuev, K D. Ridge

Abstract

Signal transduction in vertebrate photoreceptor cells begins with absorption of light by the visual pigment rhodopsin and culminates in the closure of ion channels on the plasma membrane. Consecutive binding and hydrolysis of several guanine nucleotides support the flow of information between these two events. The high levels of GTP required for G-protein activation and cGMP synthesis in the retina are likely to be supported by nucleoside diphosphate kinase (NDP-kinase). NDP-kinase presumably constitutes an integral part of the cGMP cycle (cGMP > 5'-GMP > GDP > > GTP > cGMP) and catalyzes the phosphorylation of nucleoside diphosphates to nucleoside triphosphates by a ping-pong mechanism involving a high-energy phosphorylated enzyme intermediate. The purpose of this chapter is highlight the role of NDP-kinase in visual transduction and to provide systematic approaches for the study of its biochemical properties. The focus is on the purification of the enzyme from bovine retina, its functional characterization, the cloning of two distinct isoforms from a bovine retinal library, and their heterologous expression of E.coli.

Citation

Methods In Enzymology

Volume

316

Pub Type

Journals

Keywords

enzyme, nucleotide synthesis, phototransduction, protein purification, vision

Citation

Abdulaev, N. , Kakuev, D. and Ridge, K. (2000), Bovine Retinal Nucleoside Diphosphate Kinase: Biochemistry and Molecular Cloning, Methods In Enzymology (Accessed May 10, 2024)