Molecular mechanism for the action of the anti-CD44 monoclonal antibody MEM-85
Language English Country United States Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
26066970
DOI
10.1016/j.jsb.2015.06.005
PII: S1047-8477(15)30009-5
Knihovny.cz E-resources
- Keywords
- CD44, Epitope mapping, MEM-85, Monoclonal antibody, NMR, SAXS,
- MeSH
- Hyaluronan Receptors chemistry MeSH
- Jurkat Cells MeSH
- Hyaluronic Acid chemistry MeSH
- Humans MeSH
- Epitope Mapping MeSH
- Models, Molecular MeSH
- Antibodies, Monoclonal chemistry MeSH
- Mutation MeSH
- Nuclear Magnetic Resonance, Biomolecular MeSH
- Protein Structure, Tertiary MeSH
- Binding Sites MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Hyaluronan Receptors MeSH
- Hyaluronic Acid MeSH
- Antibodies, Monoclonal MeSH
The hyaluronate receptor CD44 plays role in cell adhesion and migration and is involved in tumor metastasis. The extracellular domain of CD44 comprises the hyaluronate-binding domain (HABD) and the membrane-proximal stem region; the short intracellular portion interacts with adaptor proteins and triggers signaling pathways. Binding of hyaluronate to CD44 HABD induces an allosteric conformational change, which results in CD44 shedding. A poorly characterized epitope in human CD44 HABD is recognized by the murine monoclonal antibody MEM-85, which cross-blocks hyaluronate binding to CD44 and also induces CD44 shedding. MEM-85 is of therapeutic interest, as it inhibits growth of lung cancer cells in murine models. In this work, we employed a combination of biophysical methods to determine the MEM-85 binding epitope in CD44 HABD and to provide detailed insight into the mechanism of MEM-85 action. In particular, we constructed a single-chain variable fragment (scFv) of MEM-85 as a tool for detailed characterization of the CD44 HABD-antibody complex and identified residues within CD44 HABD involved in the interaction with scFv MEM-85 by NMR spectroscopy and mutational analysis. In addition, we built a rigid body model of the CD44 HABD-scFv MEM-85 complex using a low-resolution structure obtained by small-angle X-ray scattering. The MEM-85 epitope is situated in the C-terminal part of CD44 HABD, rather than the hyaluronate-binding groove, and the binding of MEM-85 induces a structural reorganization similar to that induced by hyaluronate. Therefore, the mechanism of MEM-85 cross-blocking of hyaluronate binding is likely of an allosteric, relay-like nature.
Faculty of Science Charles University Prague Albertov 6 Prague 2 128 40 Czech Republic
Institute of Microbiology AS CR v v i Videnska 1083 Prague 4 142 20 Czech Republic
Institute of Molecular Genetics AS CR v v i Videnska 1083 Prague 4 142 20 Czech Republic
References provided by Crossref.org
N-Glycosylation can selectively block or foster different receptor-ligand binding modes
Atomistic fingerprint of hyaluronan-CD44 binding
