Cysteine misincorporation in bacterially expressed human alpha-synuclein

FEBS Letters
Masami MasudaMasato Hasegawa

Abstract

Bacterially expressed human alpha-synuclein (alpha-syn) has been widely used in structural and functional studies. Here we show that approximately 20% of human alpha-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 alpha-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.

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Citations

Apr 16, 2011·Journal of the American Chemical Society·Eva SevcsikElizabeth Rhoades
Jan 24, 2009·The Journal of Biological Chemistry·Motokuni YonetaniMasato Hasegawa
Mar 8, 2013·Brain : a Journal of Neurology·Masami Masuda-SuzukakeMasato Hasegawa
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Aug 26, 2021·Biochemistry·Ajay Singh SawnerSamir K Maji
May 10, 2006·Biochemistry·Masami MasudaMasato Hasegawa

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