title: High concentration of carboxypeptidase M in lungs: presence of the enzyme in alveolar type I cells.
abstract: The presence of high concentrations of membrane-bound carboxypeptidase M in human, baboon, dog, and rat lung was established by employing a variety of techniques. The activity of the enzyme in the membrane-enriched fractions of human, baboon, dog, and rat lung, measured with fluorescent dansyl substrate (DNS-Ala-Arg), was 198, 261, 484, and 153 nmol/h/mg protein, respectively. This activity in the lung was much higher than that found in the heart, liver, or kidney. The enzyme, optimally active around neutral pH, was completely inhibited by 10 microM 2-mercaptomethyl-3-guanidinoethylthiopropanoic acid and was activated by 1 mM CoCl2 to 170%. Antibody to human carboxypeptidase M immunoprecipitated the solubilized carboxypeptidase from human (98%), baboon (81%), and dog (88%) lung membrane fractions. Carboxypeptidase M is attached to lung membranes by a phosphatidylinositol glycan anchor; thus, it is released with bacterial phospholipase C. Membrane fractions from cultured human pulmonary arterial endothelial cells also contained high carboxypeptidase M activity (254 nmol/h/mg protein). A Northern blot of poly(A)+ RNA from various human tissues showed the presence of a high level of carboxypeptidase M mRNA in human lung and placenta. Finally, immunohistochemistry, employing purified antibody to the enzyme, revealed in fluorescent light microscopy that carboxypeptidase M is present in alveolar type I pneumocytes and in macrophages in apparently lower concentration. In contrast, type II alveolar epithelial cells gave negative results. Because carboxypeptidase M cleaves a variety of active peptides (e.g., bradykinin, anaphylatoxins), it may protect the alveolar surface from the effects of these peptides. In addition, carboxypeptidase M could be a marker enzyme for type I cells.
authors.affiliation: Department of Pharmacology, University of Illinois, College of Medicine, Chicago 60612.
authors.forename: A
authors.initials: A
authors.lastname: Nagae
authors.forename: M
authors.initials: M
authors.lastname: Abe
authors.forename: R P
authors.initials: RP
authors.lastname: Becker
authors.forename: P A
authors.initials: PA
authors.lastname: Deddish
authors.forename: R A
authors.initials: RA
authors.lastname: Skidgel
authors.forename: E G
authors.initials: EG
authors.lastname: Erdös
mesh_terms: D015097:3-Mercaptopropionic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D000328:Adult; D000818:Animals; D015152:Blotting, Northern; D002462:Cell Membrane / Q000201:enzymology; D002478:Cells, Cultured; D003035:Cobalt / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000201:enzymology; D058851:GPI-Linked Proteins; D006168:Guinea Pigs; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007150:Immunohistochemistry; D008297:Male; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011233:Precipitin Tests; D011650:Pulmonary Alveoli / Q000166:cytology / Q000201:enzymology*; D051381:Rats; D013045:Species Specificity; D013552:Swine
chemical_list: D058851:GPI-Linked Proteins; D003035:Cobalt; C029729:2-mercaptomethyl-3-guanidinoethylthiopropionic acid; D015097:3-Mercaptopropionic Acid; C058479:carboxypeptidase M; D008666:Metalloendopeptidases; C018021:cobaltous chloride