Cysteine misincorporation in bacterially expressed human α-synuclein

被引：68

作者：

Masuda, M

Dohmae, N

Nonaka, T

Oikawa, T

Hisanaga, SI

Goedert, M

Hasegawa, M

机构：

[1] Tokyo Inst Psychiat, Dept Mol Neurobiol, Setagaya Ku, Tokyo 1568585, Japan

[2] Tokyo Metropolitan Univ, Grad Sch Sci, Mol Neurosci Lab, Tokyo 1920397, Japan

[3] RIKEN, Biomol Characterizat, Wako, Saitama 3510198, Japan

[4] Med Res Council Lab Mol Biol, Cambridge CB2 2QH, England

来源：

FEBS LETTERS | 2006年 / 580卷 / 07期

基金：

英国医学研究理事会;

关键词：

alpha-synuclein; mistranslation; cysteine; dimerization; aggregation;

D O I：

10.1016/j.febslet.2006.02.032

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：1775 / 1779

页数：5

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