Isolation and characterization of a glycine-extended form of calcitonin receptor-stimulating peptide-1: another biologically active form of calcitonin receptor-stimulating peptide-1.
In this study, we isolated a peptide eliciting a potent stimulatory effect on cAMP production in LLC-PK(1) cells from acid extracts of porcine brain. By structural analysis, this peptide was determined to be a C-terminal glycine-extended form of calcitonin receptor-stimulating peptide-1 (CRSP-1-Gly). Synthetic CRSP-1-Gly enhanced the cAMP production in COS-7 cells expressing calcitonin (CT) receptor as strongly as CRSP-1. Measurement of immunoreactive (IR) CRSP-1-Gly by radioimmunoassay using the specific antisera against CRSP-1-Gly showed that a relatively high level (>1pmol/g wet weight) of IR-CRSP-1-Gly was detected in the midbrain, hypothalamus, anterior and posterior lobes of pituitary, and thyroid gland, and the ratio of IR-CRSP-1-Gly to total IR-CRSP-1 varies from 0.02 to 0.35 in each tissue. These results suggest that CRSP-1-Gly is actually present in the tissues as one of major endogenous molecular forms of CRSP-1, and can regulate the cells expressing the CT receptor both in the central nervous system and peripheral tissues in a manner similar to that of CRSP-1. IR-CRSP-2 and IR-CRSP-3 are also present in the brain and other tissues, but their tissue concentrations are 33% on average and less than 3% that of total IR-CRSP-1, respectively.
Katafuchi
Takeshi
T
Department of Pharmacology, National Cardiovascular Center Research Institute, 5-7-1 Fujishirodai, Suita, Osaka 565-8565, Japan.
Hamano
Kazumasa
K
Kikumoto
Katsuro
K
Minamino
Naoto
N
D000818:Animals; D001923:Brain Chemistry / Q000502:physiology*; D019556:COS Cells; D002522:Chlorocebus aethiops; D000242:Cyclic AMP / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D020033:Protein Isoforms / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D018003:Receptors, Calcitonin / Q000819:agonists* / Q000378:metabolism; D013552:Swine / Q000378:metabolism*
D009419:Nerve Tissue Proteins; D020033:Protein Isoforms; D018003:Receptors, Calcitonin; D000242:Cyclic AMP