Mutations in the B1 domain of protein G that delay the onset of amyloid fibril formation in vitro

Protein Sci. 2003 Mar;12(3):567-76. doi: 10.1110/ps.0227403.

Abstract

We previously reported that under certain experimental conditions, many variants of the B1 domain of IgG-binding protein G from Streptococcus form fibrils reproducibly. The variant I6T53 was the focus of the present study because the lag phase in the kinetics of fibril formation by this variant is significantly longer than that of other variants. This lag phase is distinguished by changes in both intrinsic fluorescence intensity and in light scattering of the protein. NMR diffusion measurements suggest that the soluble protein during the lag phase is monomeric. The kinetic profiles of fibril formation are found to depend on experimental conditions. The first kinetic phase diminishes almost completely when the reaction is seeded with preformed amyloid fibrils.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amyloid / genetics
  • Amyloid / metabolism*
  • Amyloid / ultrastructure
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / pharmacology
  • Benzothiazoles
  • Fluorescence
  • Humans
  • In Vitro Techniques
  • Magnetic Resonance Spectroscopy
  • Mass Spectrometry
  • Microscopy, Electron
  • Mutation*
  • Streptococcus / chemistry
  • Thiazoles / metabolism

Substances

  • Amyloid
  • Bacterial Proteins
  • Benzothiazoles
  • IgG Fc-binding protein, Streptococcus
  • Thiazoles
  • thioflavin T