Covalent labeling in combination with mass spectrometry is a powerful approach used in structural biology to study protein structures, interactions, and dynamics. Recently, the toolbox of covalent labeling techniques has been expanded with fast fluoroalkylation of proteins (FFAP). FFAP is a novel radical labeling method that utilizes fluoroalkyl radicals generated from hypervalent Togni reagents for targeting aromatic residues. This report further demonstrates the benefits of FFAP as a new method for structural characterization of therapeutic antibodies and interaction interfaces of antigen-antibody complexes. The results obtained from human trastuzumab and its complex with human epidermal growth factor receptor 2 (HER2) correlate well with previously published structural data and demonstrate the potential of FFAP in structural biology.
- MeSH
- alkylace MeSH
- footprinting proteinů metody MeSH
- halogenace MeSH
- imunokomplex chemie MeSH
- lidé MeSH
- mapování epitopu * metody MeSH
- monoklonální protilátky chemie imunologie MeSH
- receptor erbB-2 * chemie imunologie MeSH
- trastuzumab * chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The structure of the Fc fragment of monoclonal antibody IgG2b from hybridom M75 of Mus musculus has been determined by single crystal X-ray diffraction. This is the first report of the structure of the murine immunoglobulin isotype IgG2b. The structure refined at 2.1 A resolution provides more detailed structural information about native oligosaccharides than was previously available. High-quality Fourier maps provide a clear identification of alpha-l-fucose with partial occupancy in the first branch of the antennary oligosaccharides. A unique Fc:Fc interaction was observed at the C(H)2-C(H)3 interface.
- MeSH
- glykosylace MeSH
- imunoglobulin G chemie MeSH
- imunoglobuliny - Fc fragmenty chemie MeSH
- imunokomplex chemie MeSH
- krystalizace MeSH
- krystalografie rentgenová metody MeSH
- monoklonální protilátky chemie MeSH
- myši MeSH
- oligosacharidy chemie MeSH
- sekundární struktura proteinů MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- imunoglobulin G MeSH
- imunoglobuliny - Fc fragmenty MeSH
- imunokomplex MeSH
- monoklonální protilátky MeSH
- oligosacharidy MeSH
- MeSH
- biofyzika MeSH
- biofyzikální jevy MeSH
- epitopy chemie genetika MeSH
- HIV antigeny chemie genetika MeSH
- HIV protilátky chemie farmakologie MeSH
- HIV-1 enzymologie genetika imunologie MeSH
- HIV-proteasa genetika imunologie MeSH
- imunokomplex chemie MeSH
- inhibitory HIV-proteasy chemie farmakologie MeSH
- krystalografie rentgenová MeSH
- monoklonální protilátky chemie farmakologie MeSH
- myši MeSH
- sekvence aminokyselin MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- epitopy MeSH
- HIV antigeny MeSH
- HIV protilátky MeSH
- HIV-proteasa MeSH
- imunokomplex MeSH
- inhibitory HIV-proteasy MeSH
- monoklonální protilátky MeSH
