Catecholamine‐Metabolizing Enzymes of Bovine Brain Microvessel Endothelial Cell Monolayers

Abstract

The activities of monoamine oxidase (MAO), catechol-O-methyltransferase (COMT), phenol sulfotransferase (PST), alkaline phosphatase (AP), γ-glutamyl transpeptidase (GT), and angiotensin converting enzyme (ACE) were quantitated in primary cultures of bovine brain microvessel endothelial cell monolayers and cerebral gray matter. Significant MAO-A and -B, cytosolic and membrane-bound COMT, PST, AP, GT, and ACE activities are demonstrated in bovine gray matter. By comparison, enzyme activities of the monolayers vary with the age of the monolayer and are generally higher in complete monolayers. Relative to gray matter enzyme activities, the monolayers are enriched with AP, GT, and ACE, enzymes considered to be markers for brain endothelium. Results also indicate that the activities of MAO-A and PST in the monolayers approach those found in the gray matter. Conversely, cytosolic COMT and MAO-B activities in the monolayers are negligible and much lower, respectively, compared to activities in gray matter. Additional studies with both tissues suggest that the PST of both tissues is the thermostable form of the enzyme.