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Differential epitope masking reveals synapse-specific complexes of TRPM1

Published online by Cambridge University Press:  26 January 2018

MELINA A. AGOSTO*
Affiliation:
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas 77030
IVAN A. ANASTASSOV
Affiliation:
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas 77030
THEODORE G. WENSEL
Affiliation:
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas 77030
*
*Address correspondence to: Melina A. Agosto, Department of Biochemistry and Molecular Biology, BCM-125, Baylor College of Medicine, 1 Baylor Plaza, Houston, TX 77030. E-mail: [email protected]

Abstract

The transient receptor potential channel TRPM1 is required for synaptic transmission between photoreceptors and the ON subtype of bipolar cells (ON-BPC), mediating depolarization in response to light. TRPM1 is present in the somas and postsynaptic dendritic tips of ON-BPCs. Monoclonal antibodies generated against full-length TRPM1 were found to have differential labeling patterns when used to immunostain the mouse retina, with some yielding reduced labeling of dendritic tips relative to the labeling of cell bodies. Epitope mapping revealed that those antibodies that poorly label the dendritic tips share a binding site (N2d) in the N-terminal arm near the transmembrane domain. A major splice variant of TRPM1 lacking exon 19 does not contain the N2d binding site, but quantitative immunoblotting revealed no enrichment of this variant in synaptsomes. One explanation of the differential labeling is masking of the N2d epitope by formation of a synapse-specific multiprotein complex. Identifying the binding partners that are specific for the fraction of TRPM1 present at the synapses is an ongoing challenge for understanding TRPM1 function.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2018 

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Footnotes

Current address: Department of Ophthalmology, University of California, San Francisco, California 94143

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