EF-Tu binding peptides identified, dissected, and affinity optimized by phage display.
The highly abundant GTP binding protein elongation factor Tu (EF-Tu) fulfills multiple roles in bacterial protein biosynthesis. Phage-displayed peptides with high affinity for EF-Tu were selected from a library of approximately 4.7 x 10(11) different peptides. The lack of sequence homology among the identified EF-Tu ligands demonstrates promiscuous peptide binding by EF-Tu. Homolog shotgun scanning of an EF-Tu ligand was used to dissect peptide molecular recognition by EF-Tu. All homolog shotgun scanning selectants bound to EF-Tu with higher affinity than the starting ligand. Thus, homolog shotgun scanning can simultaneously optimize binding affinity and rapidly provide detailed structure activity relationships for multiple side chains of a polypeptide ligand. The reported peptide ligands do not compete for binding to EF-Tu with various antibiotic EF-Tu inhibitors, and could identify an EF-Tu peptide binding site distinct from the antibiotic inhibitory sites.
Murase
Katsuyuki
K
Department of Chemistry, 346-D Med Sci I, University of California, Irvine, Irvine, CA 92697, USA.
Morrison
Kim L
KL
Tam
Phillip Y
PY
Stafford
Ryan L
RL
Jurnak
Frances
F
Weiss
Gregory A
GA
D000595:Amino Acid Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000378:metabolism; D006153:Guanosine Diphosphate / Q000378:metabolism; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry; D010444:Peptide Elongation Factor Tu / Q000037:antagonists & inhibitors* / Q000378:metabolism; D019151:Peptide Library; D010455:Peptides / Q000138:chemical synthesis / Q000378:metabolism* / Q000494:pharmacology*; D011487:Protein Conformation; D020133:Reverse Transcriptase Polymerase Chain Reaction
D009841:Oligonucleotides; D019151:Peptide Library; D010455:Peptides; D006153:Guanosine Diphosphate; D010444:Peptide Elongation Factor Tu