Isolation and characterization of ornitho-kininogen.
Ornitho-kininogen was purified from chicken blood plasma by a two-stage method using chromatography on columns of S-alkylated papain-Cellulofine and DEAE-5PW. The yield was 1.7 mg from 44 ml plasma. The isolated preparation gave a single band on sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS-PAGE) with or without 2-mercaptoethanol and on disc/polyacrylamide gel electrophoresis. The relative molecular mass, Mr, of ornitho-kininogen was estimated as 74,000 on SDS-PAGE using the Ferguson plot method. Ornitho-kininogen was found to have the similar properties to those of mammalian high-Mr kininogen, in terms of the amino acid composition, molecular mass, and susceptibility to plasma kallikrein. No kininogen corresponding to mammalian low-Mr kininogen and rat T-kininogen could be detected in chicken plasma. In fact, ornitho-kininogen was degraded rapidly by bovine plasma kallikrein, liberating a kinin. This kinin was isolated from the digest by reversed-phase HPLC. The primary structure of the isolated kinin was determined as Arg1-Pro2-Pro3-Gly4-Phe5-Thr6-Pro7-Leu8-Arg9. The sequence of this peptide, named ornitho-kinin, was similar to that of bradykinin except for the substitution of Thr6 and Leu8 for Ser6 and Phe8. The isolated ornitho-kinin induced a contraction of chicken smooth muscle and had a strong hypotensive effect in the chicken. However, it did not contract the isolated rat uterus. It is suggested that this specificity difference is due to the replacement of Phe8 by Leu8. The sequence of residues 1-30 of ornitho-kininogen exhibited 43% identity with that of bovine kininogen.
Kimura
M
M
Department of Biology, Faculty of Science, Kyushu University, Fukuoka, Japan.
Sueyoshi
T
T
Takada
K
K
Tanaka
K
K
Morita
T
T
Iwanaga
S
S
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000097:blood; D000818:Animals; D001794:Blood Pressure / Q000187:drug effects; D001920:Bradykinin / Q000097:blood; D002645:Chickens; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D007704:Kininogens / Q000097:blood* / Q000494:pharmacology / Q000502:physiology; D007705:Kinins / Q000097:blood; D009119:Muscle Contraction / Q000187:drug effects; D009130:Muscle, Smooth / Q000187:drug effects; D051381:Rats; D014590:Uterine Contraction / Q000187:drug effects
D000596:Amino Acids; D007704:Kininogens; D007705:Kinins; D001920:Bradykinin