Covalent cross-linking within supramolecular peptide structures

Anal Chem. 2012 Aug 7;84(15):6790-7. doi: 10.1021/ac301198c. Epub 2012 Jul 19.

Abstract

β-Sheet peptide nanostructures (e.g., amyloid fibrils) are recognized as important entities in biological systems and as functional materials in their own right. Their unique physical properties and architectural complexity, however, present a challenge for structure determination at atomic resolution. Covalent cross-linking and mass spectrometry are appealing methods for this endeavor because, potentially, a large amount of information can be extracted from a small sample in a single experiment. Previously, we described preliminary studies on the use of a photoreactive diazirine-containing amino acid to cross-link peptide monomers in nanostructures, together with the integrated separation and analysis of the products using ion mobility spectrometry coupled to conventional mass spectrometry. Here, a pH-switchable system (Aβ(16-22), a sequence from the amyloid-β peptide) was used to examine cross-linking chemistry in morphologically distinct supramolecular structures containing, or entirely composed of, diazirine-functionalized peptides. We examine the relationship between cross-linker chemistry, covalent cross-links (identified using chemical derivatization and tandem mass spectrometry), and noncovalent structure, and report differences in the site of cross-linking that can only be explained by supramolecular templating. The results demonstrate the applicability of the approach for obtaining structural restraints in ordered supramolecular assemblies, provided that a considered evaluation of the cross-linked products is undertaken.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Amyloid beta-Peptides / chemistry*
  • Cross-Linking Reagents / chemistry*
  • Diazomethane / chemistry
  • Nanostructures / chemistry
  • Peptide Fragments / chemistry*
  • Protein Structure, Secondary
  • Tandem Mass Spectrometry*

Substances

  • Amyloid beta-Peptides
  • Cross-Linking Reagents
  • Peptide Fragments
  • amyloid beta-protein (16-22)
  • Diazomethane