Four-base codon-mediated incorporation of nonnatural amino acids into proteins in a eukaryotic cell-free translation system

2005 ◽  
Vol 99 (5) ◽  
pp. 473-476 ◽  
Author(s):  
Hikaru Taira ◽  
Masaharu Fukushima ◽  
Takahiro Hohsaka ◽  
Masahiko Sisido
Keyword(s):  
Amino Acids ◽  
Eukaryotic Cell ◽  
Translation System ◽  
Nonnatural Amino Acids ◽  
Free Translation

scholarly journals Chemical aminoacylation of tRNAs with fluorinated amino acids for in vitro protein mutagenesis

2010 ◽  
Vol 6 ◽  
Author(s):  
Shijie Ye ◽  
Allison Ann Berger ◽  
Dominique Petzold ◽  
Oliver Reimann ◽  
Benjamin Matt ◽  
...  
Keyword(s):  
Amino Acids ◽  
Translation System ◽  
Biologically Relevant ◽  
Fluorinated Amino Acids ◽  
Free Translation ◽  
Trna Species ◽  
A Cell ◽  
Protein Environment ◽  
Vitro Protein

This article describes the chemical aminoacylation of the yeast phenylalanine suppressor tRNA with a series of amino acids bearing fluorinated side chains via the hybrid dinucleotide pdCpA and ligation to the corresponding truncated tRNA species. Aminoacyl-tRNAs can be used to synthesize biologically relevant proteins which contain fluorinated amino acids at specific sites by means of a cell-free translation system. Such engineered proteins are expected to contribute to our understanding of discrete fluorines’ interaction with canonical amino acids in a native protein environment and to enable the design of fluorinated proteins with arbitrary desired properties.

Incorporation of Unnatural Non-α-Amino Acids into the N Terminus of Proteins in a Cell-Free Translation System

ChemBioChem ◽  
2007 ◽  
Vol 8 (14) ◽  
pp. 1650-1653 ◽  
Author(s):  
Norihito Muranaka ◽  
Masanori Miura ◽  
Hikaru Taira ◽  
Takahiro Hohsaka
Keyword(s):  
Amino Acids ◽  
Translation System ◽  
N Terminus ◽  
Free Translation ◽  
A Cell

scholarly journals Membrane assembly of the functional KcsA potassium channel in a vesicle-based eukaryotic cell-free translation system

2014 ◽  
Vol 59 ◽  
pp. 174-183 ◽  
Author(s):  
Srujan Kumar Dondapati ◽  
Mohamed Kreir ◽  
Robert B. Quast ◽  
Doreen A Wüstenhagen ◽  
Andrea Brüggemann ◽  
...  
Keyword(s):  
Potassium Channel ◽  
Eukaryotic Cell ◽  
Translation System ◽  
Membrane Assembly ◽  
Free Translation ◽  
Kcsa Potassium Channel

Incorporation of Nonnatural Amino Acids into Proteins by Using Various Four-Base Codons in anEscherichia coliin Vitro Translation System

Biochemistry ◽  
2001 ◽  
Vol 40 (37) ◽  
pp. 11060-11064 ◽  
Author(s):  
Takahiro Hohsaka ◽  
Yuki Ashizuka ◽  
Hikaru Taira ◽  
Hiroshi Murakami ◽  
Masahiko Sisido
Keyword(s):  
Amino Acids ◽  
Translation System ◽  
Nonnatural Amino Acids

scholarly journals Protein secondary structure determines the temporal relationship between folding and disulfide formation

2020 ◽  
Vol 295 (8) ◽  
pp. 2438-2448 ◽  
Author(s):  
Philip J. Robinson ◽  
Shingo Kanemura ◽  
Xiaofei Cao ◽  
Neil J. Bulleid
Keyword(s):  
Secondary Structure ◽  
Cell Biology ◽  
Disulfide Bonds ◽  
Protein Secondary Structure ◽  
Eukaryotic Cell ◽  
Translation System ◽  
Disulfide Formation ◽  
Free Translation ◽  
Cellular Context ◽  
Disintegrin Domain

How and when disulfide bonds form in proteins relative to the stage of their folding is a fundamental question in cell biology. Two models describe this relationship: the folded precursor model, in which a nascent structure forms before disulfides do, and the quasi-stochastic model, where disulfides form prior to folding. Here we investigated oxidative folding of three structurally diverse substrates, β2-microglobulin, prolactin, and the disintegrin domain of ADAM metallopeptidase domain 10 (ADAM10), to understand how these mechanisms apply in a cellular context. We used a eukaryotic cell-free translation system in which we could identify disulfide isomers in stalled translation intermediates to characterize the timing of disulfide formation relative to translocation into the endoplasmic reticulum and the presence of non-native disulfides. Our results indicate that in a domain lacking secondary structure, disulfides form before conformational folding through a process prone to nonnative disulfide formation, whereas in proteins with defined secondary structure, native disulfide formation occurs after partial folding. These findings reveal that the nascent protein structure promotes correct disulfide formation during cotranslational folding.

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