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- Title
TACR3 mutations disrupt NK3R function through distinct mechanisms in GnRH-deficient patients.
- Authors
Noel, Sekoni D.; Abreu, Ana Paula; Shuyun Xu; Muyide, Titilayo; Gianetti, Elena; Tusset, Cintia; Carroll, Jessica; Latronico, Ana Claudia; Seminara, Stephanie B.; Carroll, Rona S.; Kaiser, Ursula B.
- Abstract
Neurokinin ? (NKB) and its G-proteincoupled receptor, NK3R, have been implicated in the neuroendocrine control of GnRH release; however, litde is known about the structure-function relationship of this ligand-receptor pair. Moreover, loss-of-function NK3R mutations cause GnRH deficiency in humans. Using missense mutations in NK3R we previously identified in patients with GnRH deficiency, we demonstrate that Y256H and Y315C NK3R mutations in the fifth and sixth transmembrane domains (TM5 and TM6), resulted in reduced whole-cell (79.3±7.2%) or plasma membrane (67.3±7.3%) levels, respectively, compared with wild-type (WT) NK3R, with near complete loss of inositol phosphate (IP) signaling, implicating these domains in receptor trafficking, processing, and/or stability. We further demonstrate in a FRETbased assay that R295S NK3R, in the third intracellular loop (IL3), bound NKB but impaired dissociation of Gq-protein subunits from the receptor compared with WT NK3R, which showed a 10.0 ± 1.3% reduction in FRET ratios following ligand binding, indicating activation of Gq-protein signaling. Interestingly, R295S NK3R, identified in the heterozygous state in a GnRHdeficient patient, also interfered with dissociation of G proteins and IP signaling from wild-type NK3R, indicative of dominant-negative effects. Collectively, our data illustrate roles for TM5 and TM6 in NK3R trafficking and ligand binding and for IL3 in NK3R signaling.-- Noel, S. D., Abreu, A. P., Xu, S., Muyide, T., Gianetti, E., Tusset, C., Carroll, J., Latronico, A. C., Seminara, S. B., Carroll, R. S., Kaiser, U. B. TACR3 mutations disrupt NK3R function through distinct mechanisms in GnRH-deficient patients.
- Subjects
LUTEINIZING hormone releasing hormone receptors; TACHYKININ receptors; GENETIC mutation; NEUROENDOCRINE tumors; CELL membranes; INOSITOL phosphates
- Publication
FASEB Journal, 2014, Vol 28, Issue 4, p1924
- ISSN
0892-6638
- Publication type
Article
- DOI
10.1096/fj.13-240630