A conformation-constrained peptide library based on insect defensin A.
Here, we reported a conformation-constrained peptide library, that was constructed based on the scaffold of a 29 amino acids peptide derived from insect defensin A. The peptide scaffold was designed utilizing the InsightII molecular modeling software and then displayed on M13 filamentous bacteriophage by fusion with coat protein III. The library was constructed by randomization of seven positions located within the two loops of the peptide scaffold generating approximately 8.3 x 10(8) transformants. Sequences from 14 randomly selected phage clones indicated that the distribution of nucleotides and amino acids paralleled with the expected frequency. Screening against the target proteins: tumor necrosis factor alpha, TNF receptor 1, TNF receptor 2 and monoclonal antibody against BMP-2 showed significant enrichment in all cases. The results presented here show that the reconstructed insect defensin A domain will be a promising non-antibody protein scaffold for the presentation of a phage-displayed constrained peptide library.
Zhao
An
A
Beijing Institute of Basic Medical Sciences, P.O. Box 130 (3), Beijing 100850, PR China. xueyn@hotmail.com
Xue
Yanning
Y
Zhang
Jie
J
Gao
Bo
B
Feng
Jiannan
J
Mao
Canquan
C
Zheng
Li
L
Liu
Nongle
N
Wang
Fang
F
Wang
Huixin
H
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D000911:Antibodies, Monoclonal; D017104:Bacteriophage M13; D036022:Capsid Proteins / Q000235:genetics; D007313:Insecta / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D019151:Peptide Library; D011485:Protein Binding / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000737:chemistry* / Q000378:metabolism; D047889:Receptors, Tumor Necrosis Factor, Type II / Q000737:chemistry* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012984:Software; D014409:Tumor Necrosis Factor-alpha / Q000737:chemistry* / Q000378:metabolism; D023084:alpha-Defensins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D000911:Antibodies, Monoclonal; D036022:Capsid Proteins; D019151:Peptide Library; D047888:Receptors, Tumor Necrosis Factor, Type I; D047889:Receptors, Tumor Necrosis Factor, Type II; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; D023084:alpha-Defensins; C025668:coat protein, Bacteriophage M13