Purification and identification of [hydroxyprolyl3]bradykinin in ascitic fluid from a patient with gastric cancer.
Kinins in the ascitic fluid from a patient with gastric cancer were purified by gel filtration and reversed-phase high-performance liquid chromatography (HPLC). Two fractions (fractions I and II) showed kinin activity. Fraction I did not correspond to either bradykinin or other known kinins, whereas fraction II corresponded to bradykinin. Fraction I contained 8 amino acid residues from bradykinin minus 1 proline plus 1 additional hydroxyproline. Sequence analysis of fraction I showed that the proline at the third amino acid residue of bradykinin was replaced by hydroxyproline. The retention time of fraction I on reversed-phase HPLC was exactly the same as that of synthetic [hydroxyprolyl3]bradykinin (Arg-Pro-Hyp-Gly-Phe-Ser-Pro-Phe-Arg) and was distinguishable from des-Pro3-bradykinin. Thus, these results demonstrate for the first time the presence of [hydroxyprolyl3]bradykinin in vivo. This is also the first report of the presence of bradykinin in human tumor ascites.
Maeda
H
H
Department of Microbiology, Kumamoto University Medical School, Japan.
Matsumura
Y
Y
Kato
H
H
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001201:Ascites / Q000378:metabolism*; D001920:Bradykinin / Q000031:analogs & derivatives* / Q000302:isolation & purification; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007705:Kinins / Q000302:isolation & purification; D008969:Molecular Sequence Data; D013274:Stomach Neoplasms / Q000378:metabolism*
D000596:Amino Acids; D007705:Kinins; C021698:bradykinin, hydroxy-Pro(3)-; D001920:Bradykinin