Membrane-bound carbonic anhydrase in the heart.
Microsomal membranes from bovine heart homogenates were subfractionated by density gradient centrifugation. Fractions with high levels of a sarcolemmal (SL) marker are enriched in specific carbonic anhydrase (CA) activity up to ninefold compared with the microsomes. Fractions with high levels of a sarcoplasmic reticulum marker and a mitochondrial marker, respectively, exhibit specific CA activities that are similar to the one found in the microsomes. Determination of cytosolic markers reveals that the CA activity in the SL fraction is not due to contamination by cytosolic CA, and it is shown by Triton X-114 phase separation that the CA activity is due to an integral membrane protein. In cryosections from rabbit heart the SL region of cardiomyocytes is stained by the fluorescent CA inhibitor dansylsulfonamide. Intracellular staining occurs also, with a pattern suggesting the presence of CA associated with intracellular membranes. Although it cannot be excluded that there is a contribution by endothelial membranes, it appears likely that most CA of the heart is bound to the SL. The possible involvement of the enzyme in extracellular proton buffering is discussed.
Bruns
W
W
Zentrum Physiologie, Medizinische Hochschule Hannover, Federal Republic of Germany.
Gros
G
G
D000818:Animals; D002256:Carbonic Anhydrases / Q000378:metabolism*; D002417:Cattle; D003902:Detergents; D006651:Histocytochemistry; D066298:In Vitro Techniques; D009206:Myocardium / Q000201:enzymology*; D017830:Octoxynol; D011092:Polyethylene Glycols; D012508:Sarcolemma / Q000201:enzymology*; D012519:Sarcoplasmic Reticulum / Q000201:enzymology*
D003902:Detergents; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40; D002256:Carbonic Anhydrases