Bloody Fact:

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transcription
translation
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(not drawn to scale)
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DiStefano et al., Curr. Op. Struct. Biol., 1998,
8, 459-465
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Semi-synthesis re-assembly of fragments,
following cleavage of a protein by protease,
CNBr, etc.. Re-ligation of amide bonds can be
achieved using proteases to catalyze the reverse
reaction
Vogel et al., Chem.Biol., 1996, 3, 295-299
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Vogel et al., Chem.Biol., 1996, 3, 295-299
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Vogel et al., Chem.Biol., 1996, 3, 295-299
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Science, 1994, 266, 243-247
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Science, 1994, 266, 243-247
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Science, 1994, 266, 243-247
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Tom Muir et al
Well come back to this(expressed peptide
ligation)
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• Unnatural amino acid mutagenesis using
  biosynthetic processes.
• This approach requires
• A unique codon/tRNA pair
• Methods for charging the tRNA with the desired
  amino acid
• In vitro reaction such that all the reagents can
  be brought together for protein synthesis

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Noren et al., Science, 1989, 244, 182-188
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Noren et al., Science, 1989, 244, 182-188
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Noren et al., Science, 1989, 244, 182-188
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Noren et al., Science, 1989, 244, 182-188
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Noren et al., Science, 1989, 244, 182-188
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• Unnatural amino acid mutagenesis using
  biosynthetic processes.
• A partial list of amino acids that have been
  incorporated into proteins using acylated
  suppressor tRNAs
• Altered
• Backbone
• Sidechain pKa
• Sidechain function

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