Quantification, isolation and structural determination of bradykinin and hydroxyprolyl-bradykinin in tumor ascites - PubMed (original) (raw)

Quantification, isolation and structural determination of bradykinin and hydroxyprolyl-bradykinin in tumor ascites

Y Matsumura et al. Adv Exp Med Biol. 1989.

Abstract

The presence of kinins in ascitic tumor fluids from rodents and human patients was identified and quantified. In bioassay, kinin content was found to be 1 to 40 ng/ml, and by enzyme immunoassay, 0.6 to 2.5 ng/ml. In particular, a high kinin content, 40 ng/ml, was found in the ascites of a gastric cancer patient by bioassay. Purification of this kinin in the ascites from the gastric cancer patient was performed by ethanol precipitation, gel filtration and reversed-phase high-performance liquid chromatography (HPLC). Two peaks (peak A and peak B) showed kinin activity. Peak A did not correspond to either bradykinin or other known kinins, such as lysyl-bradykinin and T-kinin, whereas peak B corresponded to bradykinin. Peak A contained 8 amino acid residues from bradykinin minus one proline plus an additional hydroxyproline. Sequence analysis of peak A showed that the proline at the third amino acid residue of bradykinin was replaced by hydroxyproline. The retention time of peak A on reversed-phase HPLC was exactly the same as that of synthetic hydroxyprolyl3-bradykinin (Hyp3-bradykinin) but was distinguishable from des-Pro3-bradykinin. Thus, these results demonstrate for the first time the presence of Hyp3-bradykinin in mammalian system.

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