Abstract
The TRPM8 ion channel is expressed in sensory neurons and is responsible for sensing environmental cues, such as cold temperatures and chemical compounds, including menthol and icilin. The channel functional activity is regulated by various physical and chemical factors and is likely to be preconditioned by its molecular composition. Our studies indicate that the TRPM8 channel forms a structural-functional complex with the polyester poly-(R)-3-hydroxybutyrate (PHB). We identified by mass spectrometry a number of PHB-modified peptides in the Nterminus of the TRPM8 protein and in its extracellular S3-S4 linker. Removal of PHB by enzymatic hydrolysis and site-directed mutagenesis of both the serine residues that serve as covalent anchors for PHB and adjacent hydrophobic residues that interact with the methyl groups of the polymer resulted in significant inhibition of TRPM8 channel activity. Weconclude that the TRPM8 channel undergoes posttranslational modification by PHB and that this modification is required for its normal function
Original language | English |
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Pages (from-to) | 302-315 |
Number of pages | 14 |
Journal | Cell Reports |
Volume | 4 |
Issue number | 2 |
DOIs | |
Publication status | Published - Jul 25 2013 |
Bibliographical note
Funding Information:We would like to acknowledge the work of Dr. Rosseta Reusch as a pioneer in the field of protein/PHB complexes and thank her for the inspiration for these studies as well as for providing us with the antibodies against PHB. We are thankful to Dr. Robert Winkfein for providing us with the mammalian clone of PHB-depolymerase, PhaZ7. This work was supported by American Heart Association grant SDG-2640223 (to E.Z.) and National Institutes of Health grants R01GM098052 (to E.Z.) and NS055159 (to T.R.). The authors are grateful for funding support from NIH grant NS046593 (to H.L.) and the continued support of the NINDS NeuroProteomics Core Facility at UMDNJ, New Jersey Medical School. C.C., Y.Y., Y.B., W.C., and T.L. performed experiments and analyzed data; H.L. and T.R. edited the paper; A.C. provided technical advice and help with the MS data analysis; D.J. provided phaZ7 clones and PhaZ7 antiserum and edited the paper; E.P. gave technical support and conceptual advice and edited the paper; and E.Z. developed the concept, designed and performed experiments, analyzed data, and wrote the paper.
ASJC Scopus Subject Areas
- General Biochemistry,Genetics and Molecular Biology
PubMed: MeSH publication types
- Journal Article
- Research Support, N.I.H., Extramural
- Research Support, Non-U.S. Gov't