The subcellular localization of apomucin and nonreducing terminal N-acetylgalactosamine in porcine submaxillary glands.
Antibodies prepared against enzymatically deglycosylated porcine submaxillary gland mucin (apomucin), which were unreactive with native mucin and its partially deglycosylated derivatives, were used to immunolocalize apomucin in situ. Electron microscopy of sections of Lowicryl K4M-embedded tissue reacted successively with antibodies and protein A-gold complexes showed apomucin exclusively in mucous cells within the rough endoplasmic reticulum, transitional elements of the endoplasmic reticulum, and vesicles at the cis side of the Golgi apparatus. The Golgi apparatus, forming mucous droplets, and mucous droplets contained no apomucin. Although the rough endoplasmic reticulum contained most of the apomucin in mucous cells, some cisternae of the endoplasmic reticulum and the nuclear envelope were devoid of apomucin. Examination of tissue sections treated with the glycosidases used to prepare apomucin revealed immunolabel for apomucin throughout the secretory pathway. Colloidal gold coated with Helix pomatia lectin was used to detect nonreducing N-acetylgalactosamine residues. In mucin-producing cells lectin-gold was found in the mucous droplets, the forming mucous droplets, and throughout the Golgi apparatus but mostly in the cis portion of this organelle. In tissue sections reacted successively with lectin-gold and anti-apomucin/protein A-gold, both types of gold complex could be found in the cis side of the Golgi apparatus. These data indicate that the O-glycosylation of mucin is a posttranslational event that occurs in the Golgi apparatus and begins in the cis side of the Golgi apparatus.
Deschuyteneer
M
M
Department of Biochemistry, Duke University Medical Center, Durham, North Carolina 27710.
Eckhardt
A E
AE
Roth
J
J
Hill
R L
RL
D000116:Acetylgalactosamine / Q000032:analysis*; D000818:Animals; D005688:Galactosamine / Q000031:analogs & derivatives*; D005752:Gastric Mucins; D008854:Microscopy, Electron; D010455:Peptides / Q000032:analysis*; D013347:Subcellular Fractions / Q000032:analysis; D013363:Submandibular Gland / Q000032:analysis*; D013552:Swine
D005752:Gastric Mucins; D010455:Peptides; C053366:apomucin; D005688:Galactosamine; D000116:Acetylgalactosamine