Auto-induction medium containing glyphosate for high-level incorporation of unusual aromatic amino acids into proteins.
We describe the use of an auto-induction medium containing N-(phosphono-methyl)glycine (glyphosate) as a means for high-level introduction of nonstandard aromatic amino acids into a protein. We illustrate this approach by preparing maltose binding protein (MBP) wherein all eight tryptophan residues have been replaced with 6-fluorotryptophan at an incorporation level of 99.3%. Such a high level of incorporation is important for spectroscopic investigations, in particular 19F NMR, because each species' differing amino acid sequence potentially yields a different peak pattern that complicates spectral analysis.
Neerathilingam
Muniasamy
M
Biochemistry Department, University of Wisconsin-Madison, Madison, WI 53706, USA.
Markley
John L
JL
D001709:Biotechnology / Q000379:methods; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005998:Glycine / Q000031:analogs & derivatives* / Q000378:metabolism; D006540:Herbicides / Q000378:metabolism*; D058619:Maltose-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014364:Tryptophan / Q000031:analogs & derivatives* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000097797:Glyphosate
D005998:Glycine; D006540:Herbicides; D058619:Maltose-Binding Proteins; D011994:Recombinant Proteins; D014364:Tryptophan; D000097797:Glyphosate; C001244:6-fluorotryptophan