Processing of pro-islet amyloid polypeptide in the constitutive and regulated secretory pathways of beta cells

Mol Endocrinol. 2005 Aug;19(8):2154-63. doi: 10.1210/me.2004-0407. Epub 2005 Mar 31.


Islet amyloid is a pathologic characteristic of the pancreas in type 2 diabetes comprised mainly of the beta-cell peptide islet amyloid polypeptide (IAPP; amylin). We used a pulse-chase approach to investigate the kinetics of processing and secretion of the IAPP precursor, proIAPP, in beta cells. By only 20 min after synthesis, a COOH-terminally processed proIAPP intermediate (approximately 6 kDa) was already present in beta cells. Formation of this NH2-terminally extended intermediate was not prevented by arresting secretory pathway transport at the trans-Golgi network (TGN) by either brefeldin A or temperature blockade, suggesting that this initial cleavage step occurs in the TGN before entry of (pro)IAPP into granules. Mature IAPP (approximately 4 kDa) was not detected until 60 min of chase, suggesting that NH2-terminal cleavage occurs in granules. Cells chased in low glucose without Ca2+ or with diazoxide, to block regulated release, secreted both proIAPP (approximately 8 kDa) and a partially processed form (approximately 6 kDa) via the constitutive secretory pathway. Stimulation of regulated secretion resulted in secretion primarily of mature IAPP as well as low levels of both unprocessed (approximately 8 kDa) and partially processed (approximately 6 kDa) proIAPP. We conclude that normal processing of proIAPP is a two-step process initiated by cleavage at its COOH terminus (likely by prohormone convertase 1/3 in the TGN) followed by cleavage at its NH2 terminus (by prohormone convertase 2 in granules) to form IAPP. Both proIAPP and its NH2-terminally extended intermediate appear to be normal secretory products of the beta cell that can be released via either the regulated or constitutive secretory pathways.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Amyloid / metabolism*
  • Animals
  • Brefeldin A / pharmacology
  • Cell Line, Transformed
  • Furin / chemistry
  • Immunoprecipitation
  • Insulin-Secreting Cells / metabolism*
  • Kinetics
  • Mice
  • Mice, Inbred C57BL
  • Models, Biological
  • Molecular Sequence Data
  • NIH 3T3 Cells
  • Peptides / chemistry
  • Protein Binding
  • Protein Structure, Tertiary
  • Rats
  • Retroviridae / genetics
  • Sequence Homology, Amino Acid
  • Temperature
  • Time Factors
  • trans-Golgi Network / metabolism


  • Amyloid
  • Peptides
  • pro-islet amyloid polypeptide
  • Brefeldin A
  • Furin