7KBM

Reverse Transcriptase Diabody with R83C Mutation Crystallized in C2

7KBM の概要

• エントリーDOI: 10.2210/pdb7kbm/pdb
• 分子名称: Single-chain scFv (2 entities in total)
• 機能のキーワード: diabody, immunoglobin, reverse transcriptase, protein binding
• 由来する生物種: Homo sapiens (Human)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 26403.19

構造登録者

Chesterman, C.,Arnold, E. (登録日: 2020-10-02, 公開日: 2021-03-10, 最終更新日: 2024-10-09)

主引用文献

Chesterman, C.,Arnold, E.
Co-crystallization with diabodies: A case study for the introduction of synthetic symmetry.
Structure, 29:598-605.e3, 2021

PubMed Abstract: This work presents a method for introducing synthetic symmetry into protein crystallization samples using an antibody fragment termed a diabody (Dab). These Dabs contain two target binding sites, and engineered disulfide bonds have been included to modulate Dab flexibility. The impacts of Dab engineering have been observed through assessment of thermal stability, small-angle X-ray scattering, and high-resolution crystal structures. Complexes between the engineered Dabs and HIV-1 reverse transcriptase (RT) bound to a high-affinity DNA aptamer were also generated to explore the capacity of engineered Dabs to enable the crystallization of bound target proteins. This strategy increased the crystallization hit frequency obtained for RT-aptamer, and the structure of a Dab-RT-aptamer complex was determined to 3.0-Å resolution. Introduction of synthetic symmetry using a Dab could be a broadly applicable strategy, especially when monoclonal antibodies for a target have previously been identified.

PubMed: 33636101
DOI: 10.1016/j.str.2021.02.001

実験手法

X-RAY DIFFRACTION (2 Å)