The component polypeptide chains of bovine insulin nucleate or inhibit aggregation of the parent protein in a conformation-dependent manner

J Mol Biol. 2006 Jul 7;360(2):497-509. doi: 10.1016/j.jmb.2006.05.007. Epub 2006 May 17.

Abstract

Amyloid fibrils are typically rigid, unbranched structures with diameters of approximately 10 nm and lengths up to several micrometres, and are associated with more than 20 diseases including Alzheimer's disease and type II diabetes. Insulin is a small, predominantly alpha-helical protein consisting of 51 residues in two disulfide-linked polypeptide chains that readily assembles into amyloid fibrils under conditions of low pH and elevated temperature. We demonstrate here that both the A-chain and the B-chain of insulin are capable of forming amyloid fibrils in isolation under similar conditions, with fibrillar morphologies that differ from those composed of intact insulin. Both the A-chain and B-chain fibrils were found to be able to cross-seed the fibrillization of the parent protein, although these reactions were substantially less efficient than self-seeding with fibrils composed of full-length insulin. In both cases, the cross-seeded fibrils were morphologically distinct from the seeding material, but shared common characteristics with typical insulin fibrils, including a very similar helical repeat. The broader distribution of heights of the cross-seeded fibrils compared to typical insulin fibrils, however, indicates that their underlying protofilament hierarchy may be subtly different. In addition, and remarkably in view of this seeding behavior, the soluble forms of the A-chain and B-chain peptides were found to be capable of inhibiting insulin fibril formation. Studies using mass spectrometry suggest that this behavior might be attributable to complex formation between insulin and the A-chain and B-chain peptides. The finding that the same chemical form of a polypeptide chain in different physical states can either stimulate or inhibit the conversion of a protein into amyloid fibrils sheds new light on the mechanisms underlying fibril formation, fibril strain propagation and amyloid disease initiation and progression.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amyloid / ultrastructure
  • Animals
  • Cattle
  • Insulin / chemistry*
  • Peptides / chemistry*
  • Peptides / metabolism
  • Peptides / pharmacology*
  • Protein Binding / drug effects*
  • Protein Structure, Quaternary / drug effects
  • Protein Structure, Secondary / drug effects
  • Solubility
  • Spectrometry, Mass, Electrospray Ionization
  • Time Factors

Substances

  • Amyloid
  • Insulin
  • Peptides