Carboxypeptidase M in brain and peripheral nerves.
Carboxypeptidase M (CPM), a plasma membrane-bound enzyme, cleaves C-terminal basic amino acids with a neutral pH optimum. We studied its distribution in human, baboon, and dog brain and in dog peripheral nerves. Areas were dissected, homogenized, centrifuged, and assayed for activity with dansyl-Ala-Arg. The corpus callosum and the pyramidal and optic tract were especially rich in CPM, whereas basal ganglia and cortex had low activity. The identity of the basic carboxypeptidase activity with CPM was shown by similarities in subcellular localization, membrane attachment, substrate hydrolysis, inhibition by a specific basic carboxypeptidase inhibitor, and cross-reaction with anti-human CPM antiserum. This antiserum immunoprecipitated an average of 85% of the activity in human and baboon brain and approximately 66% in dog brain. CPM co-purified with myelin extracted from the brain. Consistent with results obtained in placenta and cultured kidney cells, CPM in the brain appears to be membrane-bound via a phosphatidylinositol glycan anchor. In the peripheral nerves, the specific activity in dog sciatic nerve and in vagus was high (98 and 149 nmol/h/mg of protein, respectively). In immunohistochemical studies, glia in the brain, which appear to be oligodendrocytes or astrocytes, and the outer aspects of myelin sheaths and Schwann cells in sciatic and vagus nerves were stained. We conclude that in some areas of the CNS and the PNS, CPM is closely associated with myelin and myelin-forming cells. Northern blot analysis revealed the presence of mRNA coding for CPM in the brain, showing that the enzyme is indeed synthesized there.
Nagae
A
A
Laboratory of Peptide Research, University of Illinois College of Medicine, Chicago 60612.
Deddish
P A
PA
Becker
R P
RP
Anderson
C H
CH
Abe
M
M
Tan
F
F
Skidgel
R A
RA
Erdös
E G
EG
D015097:3-Mercaptopropionic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D015152:Blotting, Northern; D001921:Brain / Q000201:enzymology*; D001923:Brain Chemistry; D003035:Cobalt / Q000494:pharmacology; D004285:Dogs; D058851:GPI-Linked Proteins; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007106:Immune Sera; D007150:Immunohistochemistry; D001121:Lysine Carboxypeptidase / Q000037:antagonists & inhibitors; D008666:Metalloendopeptidases / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D009186:Myelin Sheath / Q000201:enzymology; D010215:Papio; D010525:Peripheral Nerves / Q000201:enzymology*; D011233:Precipitin Tests; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D012583:Schwann Cells / Q000201:enzymology
D058851:GPI-Linked Proteins; D007106:Immune Sera; D012333:RNA, Messenger; D003035:Cobalt; C029729:2-mercaptomethyl-3-guanidinoethylthiopropionic acid; D015097:3-Mercaptopropionic Acid; C058479:carboxypeptidase M; D001121:Lysine Carboxypeptidase; D008666:Metalloendopeptidases; C018021:cobaltous chloride