Production of Recombinant Alpha-Synuclein: Still No Standardized Protocol in Sight

Biomolecules. 2022 Feb 18;12(2):324. doi: 10.3390/biom12020324.

Abstract

Synucleinopathies are a group of neurodegenerative diseases, characterized by the abnormal accumulation of the protein alpha-synuclein (aSyn). aSyn is an intrinsically disordered protein that can adopt different aggregation states, some of which may be associated with disease. Therefore, understanding the transitions between such aggregation states may be essential for deciphering the molecular underpinnings underlying synucleinopathies. Recombinant aSyn is routinely produced and purified from E. coli in many laboratories, and in vitro preparations of aSyn aggregated species became central for modeling neurodegeneration in cell and animal models. Thus, reproducibility and reliability of such studies largely depends on the purity and homogeneity of aSyn preparations across batches and between laboratories. A variety of different methods are in use to produce and purify aSyn, which we review in this commentary. We also show how extraction buffer composition can affect aSyn aggregation, emphasizing the importance of standardizing protocols to ensure reproducibility between different laboratories and studies, which are essential for advancing the field.

Keywords: Parkinson’s disease; aggregation; alpha-synuclein; amyloid; synucleinopathies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Recombinant Proteins
  • Reproducibility of Results
  • Synucleinopathies
  • alpha-Synuclein* / genetics
  • alpha-Synuclein* / metabolism

Substances

  • Recombinant Proteins
  • alpha-Synuclein