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Human IgA-binding Peptides Selected from Random Peptide Libraries AFFINITY MATURATION AND APPLICATION IN IGA PURIFICATION

Authors
Hatanaka, TakaakiOhzono, ShinjiPark, MiraeSakamoto, KotaroTsukamoto, ShogoSugita, RyoheiIshitobi, HiroyukiMori, ToshiyukiIto, OsamuSorajo, KoichiSugimura, KazuhisaHam, SihyunIto, Yuji
Issue Date
Dec-2012
Publisher
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
Citation
JOURNAL OF BIOLOGICAL CHEMISTRY, v.287, no.51, pp 43126 - 43136
Pages
11
Journal Title
JOURNAL OF BIOLOGICAL CHEMISTRY
Volume
287
Number
51
Start Page
43126
End Page
43136
URI
https://scholarworks.sookmyung.ac.kr/handle/2020.sw.sookmyung/11789
DOI
10.1074/jbc.M112.389742
ISSN
0021-9258
1083-351X
Abstract
Phage display system is a powerful tool to design specific ligands for target molecules. Here, we used disulfide-constrained random peptide libraries constructed with the T7 phage display system to isolate peptides specific to human IgA. The binding clones (A1-A4) isolated by biopanning exhibited clear specificity to human IgA, but the synthetic peptide derived from the A2 clone exhibited a low specificity/affinity (K-d = 1.3 mu M). Therefore, we tried to improve the peptide using a partial randomized phage display library and mutational studies on the synthetic peptides. The designed Opt-1 peptide exhibited a 39-fold higher affinity (K-d = 33 nM) than the A2 peptide. An Opt-1 peptide-conjugated column was used to purify IgA from human plasma. However, the recovered IgA fraction was contaminated with other proteins, indicating nonspecific binding. To design a peptide with increased binding specificity, we examined the structural features of Opt-1 and the Opt-1-IgA complex using all-atom molecular dynamics simulations with explicit water. The simulation results revealed that the Opt-1 peptide displayed partial helicity in the N-terminal region and possessed a hydrophobic cluster that played a significant role in tight binding with IgA-Fc. However, these hydrophobic residues of Opt-1 may contribute to nonspecific binding with other proteins. To increase binding specificity, we introduced several mutations in the hydrophobic residues of Opt-1. The resultant Opt-3 peptide exhibited high specificity and high binding affinity for IgA, leading to successful isolation of IgA without contamination.
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