5L2H

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

Summary for 5L2H

• Entry DOI: 10.2210/pdb5l2h/pdb
• Descriptor: Centyrin, GLYCEROL (3 entities in total)
• Functional Keywords: scaffold protein, fibronectin type iii, beta sandwich, de novo protein
• Biological source: synthetic construct
• Total number of polymer chains: 4
• Total formula weight: 46067.21

Authors

Cardoso, R.M.F.,Goldberg, S.D.,O Neil, K.T.,Gilliland, G.L. (deposition date: 2016-08-01, release date: 2016-11-02, Last modification date: 2024-03-06)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (1.8013 Å)