[English] 日本語
![](img/lk-miru.gif)
- SASDAG7: CD44 HABD scFv MEM-85 complex (Hyaluronate binding domain of CD44... -
+
Open data
-
Basic information
Entry | Database: SASBDB / ID: SASDAG7 |
---|---|
![]() | CD44 HABD scFv MEM-85 complex
|
Biological species | ![]() ![]() ![]() |
![]() | ![]() Title: Molecular mechanism for the action of the anti-CD44 monoclonal antibody MEM-85. Authors: Jana Škerlová / Vlastimil Král / Michael Kachala / Milan Fábry / Ladislav Bumba / Dmitri I Svergun / Zdeněk Tošner / Václav Veverka / Pavlína Řezáčová / ![]() ![]() Abstract: 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 ...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. |
![]() |
|
-
Structure visualization
Structure viewer | Molecule: ![]() ![]() |
---|
-
Downloads & links
-Models
Model #279 | ![]() Type: dummy / Software: DAMMIN / Radius of dummy atoms: 2.50 A / Symmetry: P1 / Chi-square value: 2.460 ![]() |
---|---|
Model #280 | ![]() Type: atomic / Software: SASREF / Radius of dummy atoms: 1.90 A / Chi-square value: 1.24 ![]() |
-
Sample
![]() | Name: CD44 HABD scFv MEM-85 complex / Specimen concentration: 1.30-5.00 / Entity id: 158 / 159 |
---|---|
Buffer | Name: PBS / PK: 7 / pH: 7.4 |
Entity #158 | Name: CD44 HABD / Type: protein / Description: Hyaluronate binding domain of CD44 antigen / Formula weight: 17.9 / Num. of mol.: 1 / Source: Homo sapiens Sequence: SNAASQIDLN ITCRFAGVFH VEKNGRYSIS RTEAADLCKA FNSTLPTMAQ MEKALSIGFE TCRYGFIEGH VVIPRIHPNS ICAANNTGVY ILTSNTSQYD TYCFNASAPP EEDCTSVTDL PNAFDGPITI TIVNRDGTRY VQKGEYRTNP EDIYPSNPTD DDV |
Entity #159 | Name: scFv MEM-85 / Type: protein Description: Single-chain Variable Fragment of Antibody MEM-85 Formula weight: 28.5 / Num. of mol.: 1 / Source: Mus musculus Sequence: EVQLQESGPG LVAPSQSLSI TCTVSGFSLT NYGVHWVRQP PGKGLEWLGV IWAGGSTNYN SALMSRLSIS KDNSKSQVFL KMNSLQTDDT AMYYCARDGA RAMDYWGQGT TVTVSGGGGS GGGGSGGGGS GGGGSDIVMS QSPSSLAVSV GEKVTVSCKS SQSLLYSSNQ ...Sequence: EVQLQESGPG LVAPSQSLSI TCTVSGFSLT NYGVHWVRQP PGKGLEWLGV IWAGGSTNYN SALMSRLSIS KDNSKSQVFL KMNSLQTDDT AMYYCARDGA RAMDYWGQGT TVTVSGGGGS GGGGSGGGGS GGGGSDIVMS QSPSSLAVSV GEKVTVSCKS SQSLLYSSNQ KNYLAWYQQK PGQSPKLLIS WASTRESGVP DRFTGSGSGT DFTLTISSVK AEDLAVYYCQ QSYSYPWTFG GGTKLEIKRE QKLISEEDLN GTHHHHH |
-Experimental information
Beam | Instrument name: PETRA III P12 / City: Hamburg / 国: Germany ![]() | |||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Detector | Name: Pilatus 2M | |||||||||||||||||||||||||||||||||
Scan |
| |||||||||||||||||||||||||||||||||
Distance distribution function P(R) |
| |||||||||||||||||||||||||||||||||
Result |
|