6BI2

Trastuzumab Fab D185A (Light Chain) Mutant Biotin Conjugation.

Summary for 6BI2

• Entry DOI: 10.2210/pdb6bi2/pdb
• Related: 6BHZ 6BI0
• Descriptor: Trastuzumab Anti-HER2 Fab Heavy Chain, Trastuzumab Anti-HER2 Fab Light Chain, 1,2-ETHANEDIOL, ... (5 entities in total)
• Functional Keywords: fab, immunoglobulin, immune system
• Biological source: Homo sapiens; More
• Total number of polymer chains: 4
• Total formula weight: 95744.84

Authors

DiDonato, M.,Spraggon, G. (deposition date: 2017-10-31, release date: 2018-02-14, Last modification date: 2024-10-30)

Primary citation

Pham, G.H.,Ou, W.,Bursulaya, B.,DiDonato, M.,Herath, A.,Jin, Y.,Hao, X.,Loren, J.,Spraggon, G.,Brock, A.,Uno, T.,Geierstanger, B.H.,Cellitti, S.E.
Tuning a Protein-Labeling Reaction to Achieve Highly Site Selective Lysine Conjugation.
Chembiochem, 19:799-804, 2018

PubMed Abstract: Activated esters are widely used to label proteins at lysine side chains and N termini. These reagents are useful for labeling virtually any protein, but robust reactivity toward primary amines generally precludes site-selective modification. In a unique case, fluorophenyl esters are shown to preferentially label human kappa antibodies at a single lysine (Lys188) within the light-chain constant domain. Neighboring residues His189 and Asp151 contribute to the accelerated rate of labeling at Lys188 relative to the ≈40 other lysine sites. Enriched Lys188 labeling can be enhanced from 50-70 % to >95 % by any of these approaches: lowering reaction temperature, applying flow chemistry, or mutagenesis of specific residues in the surrounding protein environment. Our results demonstrated that activated esters with fluoro-substituted aromatic leaving groups, including a fluoronaphthyl ester, can be generally useful reagents for site-selective lysine labeling of antibodies and other immunoglobulin-type proteins.

PubMed: 29388367
DOI: 10.1002/cbic.201700611

Experimental method

X-RAY DIFFRACTION (1.801 Å)