Identification of urotensin II-related peptide as the urotensin II-immunoreactive molecule in the rat brain.
Urotensin II (UII) has been reported as the most potent known vasoconstrictor. While rat and mouse orthologs of UII precursor protein have been reported, only the tentative structures of UII peptides of these animals have been demonstrated, since prepro-UII proteins lack typical processing sites for their mature peptides. In the present study, we isolated a novel peptide, UII-related peptide (URP), from the extract of the rat brain as the sole immunoreactive substance to anti-UII antibody; the amino acid sequence of the peptide was determined as ACFWKYCV. cDNAs encoding rat, mouse, and human precursor proteins for URP were cloned and revealed that the sequences of mouse and human URP peptides are the same as that for rat URP. Prepro-URP gene is expressed in several rat tissues such as those of the thymus, spleen, testis, and spinal cord, although with lower levels than the prepro-UII gene. In the human, the prepro-URP gene is expressed comparably to prepro-UII in several tissues except the spinal cord. URP was found to bind and activate the human or rat UII receptors (GPR14) and showed a hypotensive effect when administered to anesthetized rats. These results suggest that URP is the endogenous and functional ligand for UII receptor in the rat and mouse, and possibly in the human. We also describe the preparation of specific monoclonal antibodies raised against UII peptide and the establishment of a highly sensitive enzyme immunoassay system for UII peptides.
Sugo
Tsukasa
T
Discovery Research Laboratories, Pharmaceutical Research Division, Takeda Chemical Industries, Ltd., Tsukuba, Ibaraki, Japan.
Murakami
Yuko
Y
Shimomura
Yukio
Y
Harada
Mioko
M
Abe
Michiko
M
Ishibashi
Yoshihiro
Y
Kitada
Chieko
C
Miyajima
Nobuyuki
N
Suzuki
Nobuhiro
N
Mori
Masaaki
M
Fujino
Masahiko
M
D000595:Amino Acid Sequence; D000818:Animals; D001794:Blood Pressure; D001921:Brain / Q000378:metabolism*; D016466:CHO Cells; D002118:Calcium / Q000378:metabolism; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007124:Immunoenzyme Techniques; D047908:Intracellular Signaling Peptides and Proteins; D008024:Ligands; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D036361:Peptide Hormones / Q000737:chemistry* / Q000502:physiology*; D010455:Peptides / Q000737:chemistry; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D051381:Rats; D017208:Rats, Wistar; D017386:Sequence Homology, Amino Acid; D013997:Time Factors; D014018:Tissue Distribution; D014579:Urotensins / Q000378:metabolism*
D018076:DNA, Complementary; D047908:Intracellular Signaling Peptides and Proteins; D008024:Ligands; D036361:Peptide Hormones; D010455:Peptides; C479080:UTS2B protein, human; D014579:Urotensins; C479081:urotensin II-related peptide, rat; C029677:urotensin II; D002118:Calcium