Design of a binding scaffold based on variable lymphocyte receptors of jawless vertebrates by module engineering = 모듈단백질 조합에 의한 결합스캐폴드 디자인

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Title
Design of a binding scaffold based on variable lymphocyte receptors of jawless vertebrates by module engineering = 모듈단백질 조합에 의한 결합스캐폴드 디자인
Author(s)
S C Lee; K Park; J Han; J J Lee; H J Kim; S Hong; W Heu; Yu Jung Kim; J S Ha; Seung Goo Lee; H K Cheong; Y H Jeon; D Kim; H S Kim
Bibliographic Citation
Proceedings of National Academy of Sciences of United States of America, vol. 109, no. 9, pp. 3299-3304
Publication Year
2012
Abstract
Repeat proteins have recently been of great interest as potential alternatives to immunoglobulin antibodies due to their unique structural and biophysical features. We here present the development of a binding scaffold based on variable lymphocyte receptors, which are nonimmunoglobulin antibodies composed of Leucinerich repeat modules in jawless vertebrates, by module engineering. A template scaffold was first constructed by joining consensus repeat modules between the N- and C-capping motifs of variable lymphocyte receptors. The N-terminal domain of the template scaffold was redesigned based on the internalin-B cap by analyzing the modular similarity between the respective repeat units using a computational approach. The newly designed scaffold, termed "Repebody," showed a high level of soluble expression in bacteria, displaying high thermodynamic and pH stabilities. Ease of molecular engineering was shown by designing repebodies specific for myeloid differentiation protein-2 and hen egg lysozyme, respectively, by a rational approach. The crystal structures of designed repebodies were determined to elucidate the structural features and interaction interfaces. We demonstrate general applicability of the scaffold by selecting repebodies with different binding affinities for interleukin-6 using phage display.
Keyword
Modular architectureMolecular binderNon-antibody scaffoldRepeat protein
ISSN
0027-8424
Publisher
Natl Acad Sciences
DOI
http://dx.doi.org/10.1073/pnas.1113193109
Type
Article
Appears in Collections:
Synthetic Biology and Bioengineering Research Institute > 1. Journal Articles
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