Preparation and Screening of Catalytic Amyloid Assemblies

Methods Mol Biol. 2018:1777:261-270. doi: 10.1007/978-1-4939-7811-3_16.

Abstract

Aggregation of proteins into amyloids has long been recognized as one of the major contributors to disease and aging. Amyloids are known to catalyze their own formation but they have been considered the rock-bottom thermodynamic minimum of the protein fold without much functionality. We have recently demonstrated that aggregation of short peptides in the presence of metal ions gives rise to efficient catalytic activity. Here we present a detailed protocol for the synthesis and purification of these peptides and the preparation of amyloid-like fibrils. Then we describe an easy-to-perform, high-throughput assay to measure their hydrolytic activity.

Keywords: Amyloids; Catalysis; Hydrolysis; Peptides; Self-assembly.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acids
  • Amyloid / chemical synthesis
  • Amyloid / chemistry*
  • Amyloid / metabolism
  • Amyloid beta-Peptides / chemical synthesis
  • Amyloid beta-Peptides / chemistry
  • Amyloid beta-Peptides / isolation & purification
  • Amyloid beta-Peptides / metabolism
  • Catalysis
  • Hydrogen-Ion Concentration
  • Hydrolysis
  • Peptides / chemical synthesis
  • Peptides / chemistry
  • Peptides / isolation & purification
  • Peptides / metabolism
  • Protein Aggregates*
  • Protein Multimerization*
  • Solid-Phase Synthesis Techniques

Substances

  • Amino Acids
  • Amyloid
  • Amyloid beta-Peptides
  • Peptides
  • Protein Aggregates