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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Orci, L. Amherdt, M. Ravazzola, M. Vassalli, J. D. Perrelet, A. Anderson, R. G. Madsen, O. |
| Abstract | Proinsulin is a single polypeptide chain composed of the B and A subunits of insulin joined by the C-peptide region. Proinsulin is converted to insulin during the maturation of secretory vesicles by the action of two proteases and conversion is inhibited by ionophores that disrupted intracellular H+ gradients. To determine if conversion of prohormone to hormone actually occurs in an acidic secretory vesicle, cultured rat islet cells were incubated in the presence of 3-(2,4- dinitroanilino)-3' amino-N-methyldipropylamine (DAMP), a basic congener of dinitrophenol that concentrates in acidic compartments and is retained there after aldehyde fixation. The cells were processed for indirect protein A-gold colocalization of DAMP, using a monoclonal antibody to dinitrophenol, and proinsulin, using a monoclonal antibody that exclusively reacts with the prohormone. The average density of DAMP-specific gold particles in immature secretory vesicles that contained proinsulin was 71/micron 2 (18 times cytoplasmic background), which indicated that this compartment was acidic. However, the density of DAMP-specific gold particles in the insulin-rich mature secretory vesicle averaged 433/micron 2. This suggests that although proinsulin conversion occurs in an acidic compartment, the secretory vesicles become more acidic as they mature. Since the concentration of anti- proinsulin IgG binding in secretory vesicles is inversely proportional to the conversion of proinsulin to insulin, we were able to determine that maturing secretory vesicles had to reach a critical pH before proinsulin conversion occurred. |
| ISSN | 00219525 |
| e-ISSN | 15408140 |
| Journal | The Journal of Cell Biology |
| Part | Pt 1 |
| Issue Number | 6 |
| Volume Number | 103 |
| Language | English |
| Publisher | Rockefeller University Press (United States) |
| Publisher Date | 1986-12-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Cytoplasmic Granules Metabolism Insulin Genetics Islets Of Langerhans Peptide Hydrolases Proinsulin Protein Processing, Post-Translational Animals Antibodies, Monoclonal Ultrastructure Fluorescent Antibody Technique Microscopy, Electron Rats, Inbred Strains Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Cell Biology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Medicine |
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