Salt-inducible Protein Splicing in cis and trans by Inteins from Extremely Halophilic Archaea as a Novel Protein-Engineering Tool
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- 作者：Annika Ciragan ; A. Sesilja Aranko1 ; Igor Tascon2 ; Hideo Iwaï ; ; hideo.iwai@helsinki.fi" class="auth_mail" title="E-mail the corresponding author
- 关键词：PTS ; protein trans-splicing ; GB1 ; the B1 domain of IgG binding protein G ; HSQC ; heteronuclear single quantum coherence ; TM ; transmembrane ; CTD ; C-terminal domain ; IMAC ; immobilized metal chelating ; TCEP ; tris(2-carboxyethyl)phosphine ; CPS ; conditional protein splicing ; GFP ; green fluorescent protein ; CBD ; chitin binding domain ; NOE ; nuclear Overhauser enhancement ; NOESY ; nuclear Overhauser enhancement spectroscopy ; PBS ; Phosphate-buffered saline
- 刊名：Journal of Molecular Biology
- 出版年：2016
- 出版时间：20 November 2016
- 年：2016
- 卷：428
- 期：23
- 页码：4573-4588
- 全文大小：1658 K
文摘
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Halo-obligatory inteins can be activated only above specific salinity levels.
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Salt-induced splicing was used for protein cyclization, purification, and ligation.
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Engineered MCM2 intein from H. utahensis has efficient ligation kinetics.
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Ser in proteins can be used as the + 1 position for traceless protein ligation.
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NMR study of intact TonB protein was demonstrated by segmental isotopic labeling.