carboxypeptidase-b and Hypertension

We describe the enzymes that constitute the major bradykinin (BK)-processing pathways in the perfusates of mesenteric arterial bed (MAB) and coronary vessels isolated from Wistar normotensive rats (WNR) and spontaneously hypertensive rats. The contribution of particular proteases to BK degradation was revealed by the combined analysis of fragments generated during incubation of BK with representative perfusate samples and the effect of selective inhibitors on the respective reactions. Marked differences were seen among the perfusates studied; MAB secretes, per minute of perfusion, kininase activity capable of hydrolyzing approximately 300 pmol of BK/min, which is approximately 250-fold larger amount on a per unit time basis than that of its coronary counterpart. BK degradation in the coronary perfusate seems to be mediated by ANG I-converting enzyme, neutral endopeptidase 24.11-like enzyme, and a dl-2-mercaptomethyl-3-guanidinoethylthiopropanoic acid-sensitive basic carboxypeptidase; coronary perfusate of WNR contains an additional BK-degrading enzyme whose specificity resembles that of neurolysin or thimet oligopeptidase. Diversely, a des-Arg(9)-BK-forming enzyme, responsible for nearly all of the kininase activity of MAB perfusates of WNR and spontaneously hypertensive rats, could be purified by a procedure that involved affinity chromatography over potato carboxypeptidase inhibitor-Sepharose column and shown to be structurally identical to rat pancreatic carboxypeptidase B (CPB). Comparable levels of CPB mRNA expression were observed in pancreas, liver, mesentery, and kidney, but very low levels were detected in lung, heart, aorta, and carotid artery. In conclusion, distinct BK-processing pathways operate in the perfusates of rat MAB and coronary bed, with a substantial participation of a des-Arg(9)-BK-forming enzyme identical to pancreatic CPB.

Topics: Angiotensin-Converting Enzyme Inhibitors; Animals; Blood Pressure; Bradykinin; Carboxypeptidase B; Coronary Circulation; Disease Models, Animal; Gene Expression Regulation, Enzymologic; Hydrolysis; Hypertension; Male; Metalloendopeptidases; Neprilysin; Pancreas; Peptidyl-Dipeptidase A; Perfusion; Protease Inhibitors; Rats; Rats, Inbred SHR; Rats, Wistar; Splanchnic Circulation; Substrate Specificity; Tissue Distribution

Several genetic loci involved in blood pressure regulation have recently been localized in experimental models of hypertension, but the manner in which they influence blood pressure remains unknown. Here, we report a study of the Lyon hypertensive rat strain showing that different loci are involved in the regulation of steady-state (diastolic pressure) and pulsatile (systolic-diastolic, or pulse pressure) components of blood pressure. Significant linkage was established between diastolic blood pressure and a microsatellite marker of the renin gene (REN) on rat chromosome 13, and between pulse pressure and the carboxypeptidase B gene (CPB) on chromosome 2. These findings show that two independent loci influence different haemodynamic components of blood pressure, and that pulse pressure has a specific genetic determination.

Topics: Analysis of Variance; Animals; Base Sequence; Blood Pressure; Carboxypeptidase B; Carboxypeptidases; Chromosome Mapping; Crosses, Genetic; Diastole; DNA Primers; DNA, Satellite; Female; Genetic Linkage; Genotype; Hypertension; Male; Molecular Sequence Data; Phenotype; Pulse; Rats; Rats, Inbred Strains; Renin