6AZ2

Crystal structure of Asf1-Fab 12E complex

Summary for 6AZ2

• Entry DOI: 10.2210/pdb6az2/pdb
• Related: 5uea
• Descriptor: Fab Heavy Chain, Histone chaperone ASF1, Fab Light Chain, ... (4 entities in total)
• Functional Keywords: antibody, complex, asf1, crystal chaperone, immune system
• Biological source: Homo sapiens (human); More
• Cellular location: Nucleus : P32447
• Total number of polymer chains: 6
• Total formula weight: 130587.97

Authors

Bailey, L.J.,Kossiakoff, A.A. (deposition date: 2017-09-09, release date: 2018-01-10, Last modification date: 2024-11-13)

Primary citation

Bailey, L.J.,Sheehy, K.M.,Dominik, P.K.,Liang, W.G.,Rui, H.,Clark, M.,Jaskolowski, M.,Kim, Y.,Deneka, D.,Tang, W.J.,Kossiakoff, A.A.
Locking the Elbow: Improved Antibody Fab Fragments as Chaperones for Structure Determination.
J. Mol. Biol., 430:337-347, 2018

PubMed Abstract: Antibody Fab fragments have been exploited with significant success to facilitate the structure determination of challenging macromolecules as crystallization chaperones and as molecular fiducial marks for single particle cryo-electron microscopy approaches. However, the inherent flexibility of the "elbow" regions, which link the constant and variable domains of the Fab, can introduce disorder and thus diminish their effectiveness. We have developed a phage display engineering strategy to generate synthetic Fab variants that significantly reduces elbow flexibility, while maintaining their high affinity and stability. This strategy was validated using previously recalcitrant Fab-antigen complexes where introduction of an engineered elbow region enhanced crystallization and diffraction resolution. Furthermore, incorporation of the mutations appears to be generally portable to other synthetic antibodies and may serve as a universal strategy to enhance the success rates of Fabs as structure determination chaperones.

PubMed: 29273204
DOI: 10.1016/j.jmb.2017.12.012

Experimental method

X-RAY DIFFRACTION (2.477 Å)