Structural Analysis and Optimization of Context-Independent Anti-Hypusine Antibodies

JOURNAL OF MOLECULAR BIOLOGY

卷:428

Article

Zhai, Qianting¹  He, Meng²  Song, Aimin³  Deshayes, Kurt³  Dixit, Vishva M.²  Carter, Paul J.¹ 

[1] Genentech Inc, Dept Antibody Engn, 1 DNA Way, San Francisco, CA 94080 USA

[2] Genentech Inc, Dept Physiol Chem, 1 DNA Way, San Francisco, CA 94080 USA

[3] Genentech Inc, Dept Early Discovery Biochem, 1 DNA Way, San Francisco, CA 94080 USA

关键词: hypusine;    deoxyhypusine;    eIF-5A;    antibody-peptide structure;    post-translational modification;   

DOI  :  10.1016/j.jmb.2016.01.006

来源: Elsevier

【 摘 要 】

Context-independent anti-hypusine antibodies that bind to the post-translational modification (PTM), hypusine, with minimal dependence on flanking amino acid sequences, were identified. The antibodies bind to both hypusine and deoxyhypusine or selectively to hypusine but not to deoxyhypusine. Phage display was used to further enhance the affinity of the antibodies. Affinity maturation of these anti-hypusine antibodies improved their performance in affinity capture of the only currently known hypusinated protein, eukaryotic translation initiation factor 5A. These anti-hypusine antibodies may have utility in the identification of novel hypusinated proteins. Crystal structures of the corresponding Fab fragments were determined in complex with hypusine- or deoxyhypusine-containing peptides. The hypusine or deoxyhypusine moiety was found to reside in a deep pocket formed between V-H and V-L domains of the Fab fragments. Interaction between the antibodies and hypusine includes an extensive hydrogen bond network. These are, to our knowledge, the first reported structures of context-independent anti-PTM antibodies in complex with the corresponding PTM. (C) 2016 The Authors. Published by Elsevier Ltd.

【 授权许可】

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