5L2H

Crystal Structure of W26A mutant of anti-EGFR Centyrin P54AR4-83v2

5L2H の概要

• エントリーDOI: 10.2210/pdb5l2h/pdb
• 分子名称: Centyrin, GLYCEROL (3 entities in total)
• 機能のキーワード: scaffold protein, fibronectin type iii, beta sandwich, de novo protein
• 由来する生物種: synthetic construct
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 46067.21

構造登録者

Cardoso, R.M.F.,Goldberg, S.D.,O Neil, K.T.,Gilliland, G.L. (登録日: 2016-08-01, 公開日: 2016-11-02, 最終更新日: 2024-03-06)

主引用文献

Goldberg, S.D.,Cardoso, R.M.,Lin, T.,Spinka-Doms, T.,Klein, D.,Jacobs, S.A.,Dudkin, V.,Gilliland, G.,O'Neil, K.T.
Engineering a targeted delivery platform using Centyrins.
Protein Eng. Des. Sel., 29:563-572, 2016

PubMed Abstract: Targeted delivery of therapeutic payloads to specific tissues and cell types is an important component of modern pharmaceutical development. Antibodies or other scaffold proteins can provide the cellular address for delivering a covalently linked therapeutic via specific binding to cell-surface receptors. Optimization of the conjugation site on the targeting protein, linker chemistry and intracellular trafficking pathways can all influence the efficiency of delivery and potency of the drug candidate. In this study, we describe a comprehensive engineering experiment for an EGFR binding Centyrin, a highly stable fibronectin type III (FN3) domain, wherein all possible single-cysteine replacements were evaluated for expression, purification, conjugation efficiency, retention of target binding, biophysical properties and delivery of a cytotoxic small molecule payload. Overall, 26 of the 94 positions were identified as ideal for cysteine modification, conjugation and drug delivery. Conjugation-tolerant positions were mapped onto a crystal structure of the Centyrin, providing a structural context for interpretation of the mutagenesis experiment and providing a foundation for a Centyrin-targeted delivery platform.

PubMed: 27737926
DOI: 10.1093/protein/gzw054

実験手法

X-RAY DIFFRACTION (1.8013 Å)