Rapid preparation of stable isotope labeled peptides that bind to target proteins by a phage library system.
We have developed a system for directly isolating foreign peptides displayed on the N-terminus of the major coat protein of bacteriophage M13. The phage particle in this system is formed as a mixture of wild type and modified coat proteins. The N-terminal segment of the modified coat protein was mutated for chemical cleavage, in order to obtain the displayed peptide from the major coat protein. Using 13C, 15N- labeled medium, we introduced stable isotopes, 13C and/or 15N, into the coat proteins. The NMR spectra for the cleaved peptides from the phage particles could be recorded within a few days after the selection of the phage clone.
Mizukoshi
Yumiko
Y
Japan Biological Information Research Center (JBIRC), Japan Biological Informatics Consortium (JBIC), Tokyo, 135-0064, Japan.
Takahashi
Hideo
H
Shimada
Ichio
I
D000595:Amino Acid Sequence; D017104:Bacteriophage M13 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002247:Carbon Isotopes / Q000378:metabolism; D007553:Isotope Labeling / Q000379:methods*; D008969:Molecular Sequence Data; D009587:Nitrogen Isotopes / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D036022:Capsid Proteins; D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D019151:Peptide Library; D010455:Peptides