PMID: 9438073Jan 23, 1998Paper

Functional analysis by imaging of melanophore pigment dispersion of chimeric receptors constructed by recombinant polymerase chain reaction

Brain Research. Brain Research Protocols
T S McClintock, M R Lerner

Abstract

Analysis of functional aspects of the molecular structure of proteins often requires a means to selectively alter structure and subsequently analyze function. We have adapted a method of overlap extension polymerase chain reaction (PCR) to generate multiple domain replacements in G-protein coupled receptors. The examples described herein are beta 2-adrenergic receptors whose G-protein coupling domains have been replaced by homologous domains of olfactory receptors, but the procedure has also been used to produce constructs with mutations, deletions, and fusions of two complete open reading frames. The chimeric olfactory-adrenergic receptors were assayed by functional expression in clonal lines of Xenopus melanophores. The ability of G-protein coupled second messenger pathways to cause translocation of pigment organelles within melanophores allows the use of video microscopy to assay the function of the chimeric receptors. Digital automation of microscope stage, camera, and image processing allows multiple parallel experiments to be performed. Melanophores allow responses mediated by the Gs, Gq and Gi pathways to be assayed with equal efficiency and the specificity of the coupling between chimera (or receptor) and G-protein subt...Continue Reading

References

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Mar 28, 1995·Proceedings of the National Academy of Sciences of the United States of America·J M QuillanM R Lerner
Jan 1, 1993·Ciba Foundation Symposium·M R LernerS Karne
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Citations

Oct 25, 2003·Cellular Signalling·Tony P M AnderssonSamuel P S Svensson
Mar 23, 2001·Biosensors & Bioelectronics·M F TestorfP A Oberg
Feb 19, 2002·Biosensors & Bioelectronics·Annika M KarlssonSamuel P S Svensson
Oct 16, 2003·Biosensors & Bioelectronics·Daniel FilippiniIngemar Lundström
Feb 7, 2013·Applied Neuropsychology. Adult·Kari EngRobert Denney
Jul 31, 2008·Assay and Drug Development Technologies·Anke SuskaIngemar Lundström
Jan 18, 2005·Proceedings of the National Academy of Sciences of the United States of America·Mauro Cesar IsoldiIgnacio Provencio
Jul 22, 2008·Journal of Ethnopharmacology·Therese L ErikssonRolf G G Andersson
May 20, 2003·Pigment Cell Research·Tony P M AnderssonAnnika M Karlsson
Jun 22, 2005·Biosensors & Bioelectronics·Tony P M AnderssonIngemar Lundström
Sep 8, 2009·Biosensors & Bioelectronics·Anke SuskaAnna Berghard

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