SP-Sephadex equilibrium chromatography of bradykinin and related peptides: Application to trypsin-treated human plasma |
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| Authors: | Misako U. Sampaio Marina L. Reis Edwin Fink Antonio C.M. Camargo Lewis J. Greene |
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| Affiliation: | 1. Biology Department, Brookhaven National Laboratory, Upton, New York 11973 USA;2. Protein Chemistry Laboratory, Department of Biochemistry, Faculty of Medicine of Ribeirão Preto, University of São Paulo, 14.100 Ribeirão Preto, São Paulo, Brazil;3. Department of Pharmacology, Faculty of Medicine of Ribeirão Preto, University of São Paulo, 14.100 Ribeirão Preto, São Paulo, Brazil;∗ Department of Physiological Sciences, Faculty of Dentistry and Pharmacy of Ribeirão Preto, University of São Paulo, 14.100 Ribeirão Preto, São Paulo, Brazil |
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| Abstract: | An analytical method is deseribed for the separation of bradykinin, Lys-bradykinin, and Met-Lys-bradykinin by equilibrium chromatography on SP-Sephadex C-25 eluted in 0.02 m Tris-HCl buffer, pH 8.10, 0.12 m NaCl. A second elution buffer, 0.02 m Tris-HCl buffer, pH 7.70, 0.06 m NaCl, serves as a second parameter for the identification of bradykinin and also separates the hormone from plasma bradykinin-potentiating peptides. Ten to one-hundred nanomoles of each peptide can be recovered in high yields, identified by elution position, and measured by bioassay with the isolated guinea pig ileum. The identification of bradykinin as the peptide released by trypsin acting on acid-denatured plasma is documented as an illustration of the method. |
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