Identification of a protein that confers calcitonin gene-related peptide responsiveness to oocytes by using a cystic fibrosis transmembrane conductance regulator assay.
An expression-cloning strategy was used to isolate a cDNA that encodes a protein that confers calcitonin gene-related peptide (CGRP) responsiveness to Xenopus laevis oocytes. A guinea pig organ of Corti (the mammalian hearing organ) cDNA library was screened by using an assay based on the cystic fibrosis transmembrane conductance regulator (CFTR). The CFTR is a chloride channel that is activated upon phosphorylation; this channel activity was used as a sensor for CGRP-induced activation of intracellular kinases. A cDNA library from guinea pig organ of Corti was screened by using this oocyte-CFTR assay. A cDNA was identified that contained an open reading frame coding for a small hydrophilic protein that is presumed to be either a CGRP receptor or a component of a CGRP receptor complex. This CGRP receptor component protein confers CGRP-specific activation to the CFTR assay, as no activation was detected upon application of calcitonin, amylin, neuropeptide Y, vasoactive intestinal peptide, or beta-endorphin. In situ hybridization demonstrated that the CGRP receptor component protein is expressed in outer hair cells of the organ of Corti and is colocalized with CGRP-containing efferent nerve terminals.
Luebke
A E
AE
Department of Physiology and Biophysics, Veterans Affairs Medical Center, University of Miami School of Medicine, FL 33101, USA.
Dahl
G P
GP
Roos
B A
BA
Dickerson
I M
IM
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015740:Calcitonin Gene-Related Peptide / Q000494:pharmacology*; D002712:Chlorides / Q000378:metabolism; D003001:Cloning, Molecular; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D015870:Gene Expression; D006168:Guinea Pigs; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D009865:Oocytes / Q000187:drug effects* / Q000378:metabolism*; D009925:Organ of Corti / Q000378:metabolism; D018075:RNA, Complementary / Q000235:genetics; D018015:Receptors, Calcitonin Gene-Related Peptide / Q000187:drug effects* / Q000235:genetics*; D014982:Xenopus laevis
C108477:CRCP protein, human; D002712:Chlorides; D018076:DNA, Complementary; D018075:RNA, Complementary; D018015:Receptors, Calcitonin Gene-Related Peptide; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D015740:Calcitonin Gene-Related Peptide