Cyclophilin-facilitated bradykinin inactivation in the perfused rat lung.
Cis and trans isomers of X-proline (X-Pro) bonds can influence some aspects of the kinetics of peptide metabolism. We previously used the peptidyl-prolyl cis-trans isomerase, cyclophilin, to show that angiotensin converting enzyme (ACE) preferentially hydrolyzes the trans isomer of a synthetic tripeptide that contains a C-terminal proline (Dawson et al., Am J Physiol 257:H853-H865, 1989; Merker et al., J Appl Physiol 75: 1519-1524, 1993). Bradykinin (Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg) exists as both cis and trans isomers at all three X-Pro bonds, and although its inactivation in the lung by pulmonary endothelial peptidases is extensive, commonly a small fraction of the peptide survives passage through the lung. To determine whether the presence of cis X-Pro bonds might limit the extent of bradykinin metabolism in the lung, we studied inactivation of bradykinin by the isolated perfused rat lung using the rabbit jugular vein superfused with the pulmonary venous effluent as a bioassay for bradykinin. A large fraction (> 90%) of the bradykinin in a bolus injection was inactivated in a single transit through the pulmonary circulation, but a detectable fraction emerged in the venous effluent. The addition of cyclophilin to the bradykinin in the bolus reduced the bradykinin emerging from the lungs to virtually undetectable levels. When the isomerase inhibitor cyclosporin A was included with bradykinin and cyclophilin in the injectate, this effect of cyclophilin was reversed. These observations suggest that the fraction of bradykinin that normally survives passage through the lungs contains isomers that have at least one X-Pro bond that is refractory to enzymatic inactivation and whose isomerization time constant is significantly longer than the pulmonary capillary transit time.
Merker
M P
MP
Department of Anesthesiology, Medical College of Wisconsin, Milwaukee 53226, USA.
Dawson
C A
CA
D000591:Amino Acid Isomerases / Q000494:pharmacology*; D000595:Amino Acid Sequence; D000818:Animals; D001681:Biological Assay; D001920:Bradykinin / Q000493:pharmacokinetics*; D002352:Carrier Proteins / Q000494:pharmacology*; D003524:Cyclosporins / Q000494:pharmacology*; D008168:Lung / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D019696:Peptidylprolyl Isomerase; D010477:Perfusion / Q000295:instrumentation; D051381:Rats; D017207:Rats, Sprague-Dawley; D013237:Stereoisomerism
D002352:Carrier Proteins; D003524:Cyclosporins; D000591:Amino Acid Isomerases; D019696:Peptidylprolyl Isomerase; D001920:Bradykinin