3PP4

Epitope characterization and crystal structure of GA101 provide insights into the molecular basis for the type I / type II distinction of anti- CD20 antibodies

Summary for 3PP4

• Entry DOI: 10.2210/pdb3pp4/pdb
• Related: 3PP3
• Descriptor: GA101 Fab heavy chain, GA101 Fab light chain, B-lymphocyte antigen CD20, ... (5 entities in total)
• Functional Keywords: antibody fab fragment ig-domain, cd20, cyclic peptide, antibody antigen, immune system
• Biological source: Mus musculus (mouse); More
• Cellular location: Membrane; Multi-pass membrane protein: P11836
• Total number of polymer chains: 3
• Total formula weight: 50822.21

Authors

Hopfner, K.-P.,Lammens, A. (deposition date: 2010-11-24, release date: 2011-04-13, Last modification date: 2024-10-30)

Primary citation

Niederfellner, G.,Lammens, A.,Mundigl, O.,Georges, G.J.,Schaefer, W.,Schwaiger, M.,Franke, A.,Wiechmann, K.,Jenewein, S.,Slootstra, J.W.,Timmerman, P.,Brannstrom, A.,Lindstrom, F.,Mossner, E.,Umana, P.,Hopfner, K.P.,Klein, C.
Epitope characterization and crystal structure of GA101 provide insights into the molecular basis for type I/II distinction of CD20 antibodies.
Blood, 118:358-367, 2011

PubMed Abstract: CD20 is a cell-surface marker of normal and malignant B cells. Rituximab, a monoclonal antibody targeting CD20, has improved the treatment of malignant lymphomas. Therapeutic CD20 antibodies are classified as either type I or II based on different mechanisms of killing malignant B cells. To reveal the molecular basis of this distinction, we fine-mapped the epitopes recognized by both types. We also determined the first X-ray structure of a type II antibody by crystallizing the obinutuzumab (GA101) Fab fragment alone and in complex with a CD20 cyclopeptide. Despite recognizing an overlapping epitope, GA101 binds CD20 in a completely different orientation than type I antibodies. Moreover, the elbow angle of GA101 is almost 30° wider than in type I antibodies, potentially resulting in different spatial arrangements of 2 CD20 molecules bound to a single GA101 or rituximab molecule. Using protein tomography, different CD20 complexes were found to be associated with the 2 antibodies, and confocal microscopy showed different membrane compartmentalization of these subpopulations of the cellular CD20 pool. Our findings offer a possible molecular explanation for the different cellular responses elicited by type I and II antibodies.

PubMed: 21444918
DOI: 10.1182/blood-2010-09-305847

Experimental method

X-RAY DIFFRACTION (1.6 Å)