Antibody variable domain interface and framework sequence requirements for stability and function by high-throughput experiments

Hung Ju Hsu, Kuo Hao Lee, Jhih Wei Jian, Hung Ju Chang, Chung Ming Yu, Yu Ching Lee, Ing Chien Chen, Hung Pin Peng, Chih Yuan Wu, Yu Feng Huang, Chih Yun Shao, Kuo Ping Chiu, An Suei Yang

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Protein structural stability and biological functionality are dictated by the formation of intradomain cores and interdomain interfaces, but the intricate sequence-structure-function interrelationships in the packing of protein cores and interfaces remain difficult to elucidate due to the intractability of enumerating all packing possibilities and assessing the consequences of all the variations. In this work, groups of β strand residues of model antibody variable domains were randomized with saturated mutagenesis and the functional variants were selected for high-throughput sequencing and high-throughput thermal stability measurements. The results show that the sequence preferences of the intradomain hydrophobic core residues are strikingly flexible among hydrophobic residues, implying that these residues are coupled indirectly with antigen binding through energetic stabilization of the protein structures. By contrast, the interdomain interface residues are directly coupled with antigen binding. The interdomain interface should be treated as an integral part of the antigen-binding site.

Original languageEnglish
Pages (from-to)22-34
Number of pages13
JournalStructure
Volume22
Issue number1
DOIs
Publication statusPublished - Jan 7 2014
Externally publishedYes

ASJC Scopus subject areas

  • Molecular Biology
  • Structural Biology

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    Hsu, H. J., Lee, K. H., Jian, J. W., Chang, H. J., Yu, C. M., Lee, Y. C., Chen, I. C., Peng, H. P., Wu, C. Y., Huang, Y. F., Shao, C. Y., Chiu, K. P., & Yang, A. S. (2014). Antibody variable domain interface and framework sequence requirements for stability and function by high-throughput experiments. Structure, 22(1), 22-34. https://doi.org/10.1016/j.str.2013.10.006