Cytoplasmic carbonic anhydrase II of rat coagulating gland is secreted via the apocrine export mode.
Two different pathways for protein secretion are described for epithelial cells of rat coagulating gland and dorsal prostate: the classical merocrine and the alternative apocrine release mode. Apocrine-secreted proteins are synthesized on cytoplasmic polyribosomes and are subsequently exported in protrusions on the apical cell surface (aposomes). In this article we report the identification and purification to homogeneity of a 29-kD protein from the secretion of rat coagulating gland. N-terminal amino acid sequence analyses revealed 100% identity to rat brain carbonic anhydrase II (CAH II). In addition, the 29-kD protein showed CAH enzyme activity. On Western blot analysis, a polyclonal anti-CAH II antibody raised in rabbit reacted specifically with the rat and human but not bovine CAH II isoforms. Immunohistochemical studies on rat coagulating gland showed strong labeling for CAH II protein in aposomes. Immunoelectron microscopy confined CAH II protein to the cytoplasm and aposomes, whereas no staining was visible in the compartments of the classical merocrine route, the endoplasmic reticulum and Golgi apparatus. The resident cytoplasmic protein lactate dehydrogenase, however, was not found in the secretion. Taken together, the morphological and biochemical data clearly indicate that cytoplasmic CAH II from rat coagulating gland is specifically selected and then secreted via the apocrine pathway.
Wilhelm
B
B
Department of Anatomy and Cell Biology, Philipps-University, Marburg, Germany.
Keppler
C
C
Hoffbauer
G
G
Lottspeich
F
F
Linder
D
D
Meinhardt
A
A
Aumüller
G
G
Seitz
J
J
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002256:Carbonic Anhydrases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D002417:Cattle; D003593:Cytoplasm / Q000201:enzymology* / Q000648:ultrastructure; D005089:Exocytosis; D006801:Humans; D007150:Immunohistochemistry; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008297:Male; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D011467:Prostate / Q000201:enzymology* / Q000648:ultrastructure; D051381:Rats; D017208:Rats, Wistar
D007770:L-Lactate Dehydrogenase; D002256:Carbonic Anhydrases