Book chapter
Assays of G protein/cGMP-phosphodiesterase interactions
Methods in Enzymology, pp.27-37
Elsevier Science & Technology
2002
DOI: 10.1016/S0076-6879(02)45004-5
Abstract
Activation of cGMP phosphodiesterase (PDE) by the photoreceptor G protein transducin (G1), is a key event in the vertebrate visual transduction cascade. The assays of cGMP hydrolysis by activated PDE have been a major tool for monitoring the interaction between transducin and PDE, and initial studies mainly relied on such assays. The presence of rod outer segment (ROS) membranes or lipid vesicles significantly enhances the effectiveness of PDE stimulation because of the formation of an active membrane-bound complex between G1α and PDE and allows an increase in assay sensitivity. However, membrane-supported activation of PDE by transducin is a complex process that depends on a number of factors such as type and concentration of membranes {vesicles), binding of PDE, binding of G1α to membranes, and the intact stale of lipid modifications on Pαβ and G1α. Furthermore, PDE activation is not a direct monitor of transducin binding to the enzyme. This is evident from studies of G1α mutants that bind but fail to activate PDE, or interact with the effector weakly without causing enzyme activation.
Details
- Title: Subtitle
- Assays of G protein/cGMP-phosphodiesterase interactions
- Creators
- Alexey E GranovskyNikolai O Artemyev
- Resource Type
- Book chapter
- Publication Details
- Methods in Enzymology, pp.27-37
- Publisher
- Elsevier Science & Technology
- DOI
- 10.1016/S0076-6879(02)45004-5
- eISSN
- 1557-7988
- ISSN
- 0076-6879
- Language
- English
- Date published
- 2002
- Academic Unit
- Molecular Physiology and Biophysics; Ophthalmology and Visual Sciences; Iowa Neuroscience Institute
- Record Identifier
- 9984070878102771
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