6K69

Application of anti-helix antibodies in protein structure determination (9213-3LRH)

6K69 の概要

• エントリーDOI: 10.2210/pdb6k69/pdb
• 分子名称: 3LRH intrabody, Engineered T4 lysozyme (3 entities in total)
• 機能のキーワード: antibody, protein design, structural protein
• 由来する生物種: Homo sapiens; 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 37073.46

構造登録者

Lee, J.O.,Jin, M.S.,Kim, J.W.,Kim, S.,Lee, H.,Cho, G.Y. (登録日: 2019-06-01, 公開日: 2019-08-14, 最終更新日: 2023-11-22)

主引用文献

Kim, J.W.,Kim, S.,Lee, H.,Cho, G.,Kim, S.C.,Lee, H.,Jin, M.S.,Lee, J.O.
Application of antihelix antibodies in protein structure determination.
Proc.Natl.Acad.Sci.USA, 116:17786-17791, 2019

PubMed Abstract: Antibodies are indispensable tools in protein engineering and structural biology. Antibodies suitable for structural studies should recognize the 3-dimensional (3D) conformations of target proteins. Generating such antibodies and characterizing their complexes with antigens take a significant amount of time and effort. Here, we show that we can expand the application of well-characterized antibodies by "transplanting" the epitopes that they recognize to proteins with completely different structures and sequences. Previously, several antibodies have been shown to recognize the alpha-helical conformation of antigenic peptides. We demonstrate that these antibodies can be made to bind to a variety of unrelated "off-target" proteins by modifying amino acids in the preexisting alpha helices of such proteins. Using X-ray crystallography, we determined the structures of the engineered protein-antibody complexes. All of the antibodies bound to the epitope-transplanted proteins, forming accurately predictable structures. Furthermore, we showed that binding of these antihelix antibodies to the engineered target proteins can modulate their catalytic activities by trapping them in selected functional states. Our method is simple and efficient, and it will have applications in protein X-ray crystallography, electron microscopy, and nanotechnology.

PubMed: 31371498
DOI: 10.1073/pnas.1910080116

実験手法

X-RAY DIFFRACTION (2.401 Å)