Protein semi-synthesis in living cells

Journal of the American Chemical Society

Volumn 125, Issue 24, 2003, Pages 7180-7181

  Giriat, Izabela ^a   Muir, Tom W ^a  

^a ROCKEFELLER UNIVERSITY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CELL PROTEIN; GREEN FLUORESCENT PROTEIN;

ANIMAL CELL; ARTICLE; CELL; CELLULAR DISTRIBUTION; CHEMICAL MODIFICATION; CONTROLLED STUDY; DISULFIDE BOND; GENE PROBE; GENETIC TRANSFECTION; IMMUNOPRECIPITATION; ISOTOPE LABELING; NONHUMAN; PROTEIN ANALYSIS; PROTEIN LOCALIZATION; PROTEIN PROTEIN INTERACTION; PROTEIN SYNTHESIS;

ANIMALS; CHO CELLS; CRICETINAE; DNA POLYMERASE III; GREEN FLUORESCENT PROTEINS; LUMINESCENT PROTEINS; PROTEIN BIOSYNTHESIS; PROTEIN SPLICING; PROTEIN STRUCTURE, TERTIARY; PROTEINS; TRANSDUCTION, GENETIC; TRANSFECTION;

EID: 0037774580     PISSN: 00027863     EISSN: None     Source Type: Journal    
DOI: 10.1021/ja034736i     Document Type: Article

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17. cprobe component of the system. See Supporting Information.
18. Protein splicing requires two reactive cysteines to take place. Lysis under alkylating conditions (PBS containing 1% (w/v) docecylmaltoside (DM), 10 mM iodoacetamide, 2 mM N-ethyl-maleamide, and protease inhibitors) blocks all cysteine sulhydryl groups, therefore preventing splicing.
19. c-FLAG peptide was added in large excess.
20. c(SR)-FLAG).
21. The current system has been applied successfully to the semi-synthesis of four other proteins in vivo. See Supporting Information.