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Molecular mechanism for the action of the anti-CD44 monoclonal antibody MEM-85
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SYSNO ASEP 0455882 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Molecular mechanism for the action of the anti-CD44 monoclonal antibody MEM-85 Tvůrce(i) Škerlová, Jana (UMG-J)
Král, Vlastimil (UMG-J) RID
Kachala, M. (DE)
Fábry, Milan (UMG-J) RID
Bumba, Ladislav (MBU-M) RID, ORCID
Svergun, D.I. (DE)
Tosner, Z. (CZ)
Veverka, V. (CZ)
Řezáčová, Pavlína (UMG-J) RIDZdroj.dok. Journal of Structural Biology. - : Elsevier - ISSN 1047-8477
Roč. 191, č. 2 (2015), s. 214-223Poč.str. 10 s. Jazyk dok. eng - angličtina Země vyd. US - Spojené státy americké Klíč. slova CD44 ; Epitope mapping ; Monoclonal antibody ; MEM-85 ; SAXS ; NMR Vědní obor RIV EB - Genetika a molekulární biologie CEP GA15-11851S GA ČR - Grantová agentura ČR Institucionální podpora UMG-J - RVO:68378050 ; MBU-M - RVO:61388971 UT WOS 000359096700015 DOI 10.1016/j.jsb.2015.06.005 Anotace 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 scEv 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. (C) 2015 Elsevier Inc. All rights reserved. Pracoviště Ústav molekulární genetiky Kontakt Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Rok sběru 2016
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