Localization of cellubrevin to the Golgi complex in pancreatic acinar cells.
Cellubrevin is the smallest (14 kDa) isoform of the synaptobrevin (VAMP) protein family and is found in a wide variety of tissues. Western blot analysis with a polyclonal antibody against the unique N-terminus of cellubrevin identified a protein of 14 kDa in rat pancreas. This protein distributed predominantly to the particulate fractions from the rat exocrine pancreas and was totally resistant to NaHCO3 washes, indicating that it is an integral membrane protein. Subcellular fractionation of pancreatic homogenates showed enrichment of this protein in the smooth microsomal fraction while negligible amounts were present in the zymogen granule membrane or the rough microsomal membrane fractions. As seen in other tissues, the 14 kDa immunoreactive form was proteolyzed by tetanus toxin. Light and electron microscopic immunocytochemistry localized cellubrevin immunoreactivity primarily to small vesicles and condensing vacuoles originating from the Golgi region, with significantly lower labeling on zymogen granules. Based on the intracellular localization of cellubrevin detected in acinar cells by immunocytochemistry and cell fractionation, we suggest that cellubrevin may be involved in the maturation of secretory granules.
Sengupta
D
D
Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510, USA.
Gumkowski
F D
FD
Tang
L H
LH
Chilcote
T J
TJ
Jamieson
J D
JD
D000595:Amino Acid Sequence; D000818:Animals; D002458:Cell Fractionation; D003594:Cytoplasmic Granules / Q000737:chemistry / Q000648:ultrastructure; D006056:Golgi Apparatus / Q000737:chemistry* / Q000648:ultrastructure; D015151:Immunoblotting; D007150:Immunohistochemistry; D008297:Male; D008565:Membrane Proteins / Q000032:analysis* / Q000737:chemistry / Q000187:drug effects / Q000648:ultrastructure; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D010179:Pancreas / Q000737:chemistry* / Q000648:ultrastructure; D051381:Rats; D017207:Rats, Sprague-Dawley; D013744:Tetanus Toxin / Q000494:pharmacology; D014617:Vacuoles / Q000737:chemistry / Q000648:ultrastructure; D051040:Vesicle-Associated Membrane Protein 3
D008565:Membrane Proteins; D013744:Tetanus Toxin; D051040:Vesicle-Associated Membrane Protein 3