Journal
FEBS LETTERS
Volume 580, Issue 7, Pages 1775-1779Publisher
WILEY
DOI: 10.1016/j.febslet.2006.02.032
Keywords
alpha-synuclein; mistranslation; cysteine; dimerization; aggregation
Funding
- MRC [MC_U105184291] Funding Source: UKRI
- Medical Research Council [MC_U105184291] Funding Source: Medline
- Medical Research Council [MC_U105184291] Funding Source: researchfish
Ask authors/readers for more resources
Bacterially expressed human a-synuclein (alpha-syn) has been widely used in structural and functional studies. Here we show that similar to 20% of human a-syn expressed in Escherichia coli is mistranslated and that a Cys residue is incorporated at position 136 instead of a Tyr. Site-directed mutagenesis of codon 136 (TAC to TAT) resulted in the expression of a-syn lacking Cys. Although wild-type (Y136-TAC and Y136-TAT) and mutant (C136-TGC) alpha-syn had similar propensities to assemble into filaments, the levels of dimeric alpha-syn were increased by misincorporation. To avoid potential artefacts, we recommend use of the Y136-TAT construct for the expression of human alpha-syn. (c) 2006 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available