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Title: Phosphatidylinositol 4,5-Bisphosphate Clusters the Cell Adhesion Molecule CD44 and Assembles a Specific CD44-Ezrin Heterocomplex, as Revealed by Small Angle Neutron Scattering

Journal Article · · Journal of Biological Chemistry
 [1];  [1];  [2];  [3];  [3];  [3];  [2];  [1];  [1]
  1. City College of New York, NY (United States)
  2. Icahn School of Medicine at Mount Sinai, New York, NY (United States)
  3. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

The cell adhesion molecule CD44 regulates diverse cellular functions, including cell-cell and cell-matrix interaction, cell motility, migration, differentiation, and growth. In cells, CD44 co-localizes with the membrane-cytoskeleton adapter protein Ezrin, which links the CD44 assembled receptor signaling complexes to the cytoskeletal actin and organizes the spatial and temporal localization of signaling events. Here we report that the cytoplasmic tail of CD44 (CD44ct) is largely disordered and adopts an autoinhibited conformation, which prevents CD44ct from binding directly to activated Ezrin in solution. Binding to the signaling lipid phosphatidylinositol 4,5-biphosphlate (PIP2) disrupts autoinhibition in CD44ct, and activates CD44ct to associate with Ezrin. Further, using contrast variation small angle neutron scattering, we show that PIP2 mediates the assembly of a specific hetero-tetramer complex of CD44ct with Ezrin. This study reveals a novel autoregulation mechanism in the cytoplasmic tail of CD44 and the role of PIP2 in mediating the assembly of multimeric CD44ct-Ezrin complexes. We hypothesize that polyvalent electrostatic interactions are responsible for the assembly of multimeric PIP2-CD44-Ezrin complexes.

Research Organization:
Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States). Spallation Neutron Source (SNS)
Sponsoring Organization:
USDOE Office of Science (SC)
Grant/Contract Number:
AC05-00OR22725
OSTI ID:
1185789
Journal Information:
Journal of Biological Chemistry, Vol. 290, Issue 10; ISSN 0021-9258
Publisher:
American Society for Biochemistry and Molecular BiologyCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 26 works
Citation information provided by
Web of Science

References (75)

Hyaluronan: from extracellular glue to pericellular cue journal July 2004
Using Situs for the registration of protein structures with low-resolution bead models from X-ray solution scattering journal November 2001
Essential functions of ezrin in maintenance of cell shape and lamellipodial extension in normal and transformed fibroblasts journal September 1997
Uniqueness of ab initio shape determination in small-angle scattering journal April 2003
Ezrin Controls the Macromolecular Complexes Formed between an Adapter Protein Na + /H + Exchanger Regulatory Factor and the Cystic Fibrosis Transmembrane Conductance Regulator journal August 2005
Ligand-Induced Dynamic Changes in Extended PDZ Domains from NHERF1 journal July 2013
Activation of Nanoscale Allosteric Protein Domain Motion Revealed by Neutron Spin Echo Spectroscopy journal November 2010
PTEN-Mediated Apical Segregation of Phosphoinositides Controls Epithelial Morphogenesis through Cdc42 journal January 2007
Absolute calibration of small-angle neutron scattering data journal February 1987
Ezrin self-association involves binding of an N-terminal domain to a normally masked C-terminal domain that includes the F-actin binding site. journal August 1995
A novel PKC-regulated mechanism controls CD44–ezrin association and directional cell motility journal May 2002
The use of glycerol in crystallization of T7 RNA polymerase: Implications for the use of cosolvents in crystallizing flexible proteins journal August 1990
Molecular Conformation of the Full-Length Tumor Suppressor NF2/Merlin—A Small-Angle Neutron Scattering Study journal July 2014
ERM family members as molecular linkers between the cell surface glycoprotein CD44 and actin-based cytoskeletons. journal July 1994
EPI64 regulates microvillar subdomains and structure journal December 2006
PIP 2 and Proteins: Interactions, Organization, and Information Flow journal June 2002
Structure of the Active N-terminal Domain of Ezrin: CONFORMATIONAL AND MOBILITY CHANGES IDENTIFY KEYSTONE INTERACTIONS journal November 2002
CD44 isoforms are heterogeneously expressed in breast cancer and correlate with tumor subtypes and cancer stem cell markers journal September 2011
CD44: can a cancer-initiating cell profit from an abundantly expressed molecule? journal March 2011
Spatial control of EGF receptor activation by reversible dimerization on living cells journal March 2010
ERM proteins and merlin: integrators at the cell cortex journal August 2002
Basic Motifs Target PSGL-1, CD43, and CD44 to Plasma Membrane Sites Where HIV-1 Assembles journal October 2014
NELL2-Robo3 complex structure reveals mechanisms of receptor activation for axon guidance journal March 2020
The role of the CD44 transmembrane and cytoplasmic domains in co-ordinating adhesive and signalling events journal December 2003
CD44+/CD24-breast cancer cells exhibit enhanced invasive properties: an early step necessary for metastasis journal October 2006
A Method for Determining Transmembrane Helix Association and Orientation in Detergent Micelles Using Small Angle X-Ray Scattering journal August 1999
Membrane protein sequestering by ionic protein–lipid interactions journal October 2011
Intracellular Domain Fragment of CD44 Alters CD44 Function in Chondrocytes journal July 2013
Size-Distribution Analysis of Macromolecules by Sedimentation Velocity Ultracentrifugation and Lamm Equation Modeling journal March 2000
Enhanced piezoelectricity from highly polarizable oriented amorphous fractions in biaxially oriented poly(vinylidene fluoride) with pure β crystals journal January 2021
CD44 is required for two consecutive steps in HGF/c-Met signaling journal December 2002
Using an in-vacuum CCD detector for simultaneous small- and wide-angle scattering at beamline X9 journal January 2013
The extended Q -range small-angle neutron scattering diffractometer at the SNS journal July 2010
How Random are Intrinsically Disordered Proteins? A Small Angle Scattering Perspective journal February 2012
A Conformational Switch in the Scaffolding Protein NHERF1 Controls Autoinhibition and Complex Formation journal December 2009
UCSF Chimera?A visualization system for exploratory research and analysis journal January 2004
Clustered PI(4,5)P2 accumulation and ezrin phosphorylation in response to CLIC5A journal October 2014
Structure-specific DNA-induced Conformational Changes in Taq Polymerase Revealed by Small Angle Neutron Scattering journal July 2004
Ezrin/Radixin/Moesin (ERM) Proteins Bind to a Positively Charged Amino Acid Cluster in the Juxta-Membrane Cytoplasmic Domain of CD44, CD43, and ICAM-2 journal February 1998
The role of the CD44/ezrin complex in cancer metastasis journal May 2003
Phosphoinositide binding and phosphorylation act sequentially in the activation mechanism of ezrin journal March 2004
GNOM – a program package for small-angle scattering data processing journal October 1991
Self-masking in an Intact ERM-merlin Protein: An Active Role for the Central α-Helical Domain journal February 2007
Regulation mechanism of ERM (ezrin/radixin/moesin) protein/plasma membrane association: possible involvement of phosphatidylinositol turnover and Rho-dependent signaling pathway. journal October 1996
Open Conformation of Ezrin Bound to Phosphatidylinositol 4,5-Bisphosphate and to F-actin Revealed by Neutron Scattering journal August 2012
Prospective identification of tumorigenic breast cancer cells journal March 2003
Protein Kinase C Phosphorylation Disrupts Na + /H + Exchanger Regulatory Factor 1 Autoinhibition and Promotes Cystic Fibrosis Transmembrane Conductance Regulator Macromolecular Assembly journal July 2007
Mutagenesis of the Phosphatidylinositol 4,5-Bisphosphate (Pip2) Binding Site in the Nh2-Terminal Domain of Ezrin Correlates with Its Altered Cellular Distribution journal November 2000
Determination of molecular weight by neutron scattering journal November 1981
Restoring Low Resolution Structure of Biological Macromolecules from Solution Scattering Using Simulated Annealing journal June 1999
Electrostatic Sequestration of PIP2 on Phospholipid Membranes by Basic/Aromatic Regions of Proteins journal April 2004
Regulation of Protein Activities by Phosphoinositide Phosphates journal November 2005
CD44: From adhesion molecules to signalling regulators journal January 2003
A novel mechanism of regulating breast cancer cell migration via palmitoylation-dependent alterations in the lipid raft affiliation of CD44 journal February 2014
Ezrin Induces Long-Range Interdomain Allostery in the Scaffolding Protein NHERF1 journal September 2009
Structural basis for CD44 recognition by ERM proteins journal August 2008
Biomolecular solution X-ray scattering at the National Synchrotron Light Source journal November 2010
Sedimentation equilibrium analysis of protein interactions with global implicit mass conservation constraints and systematic noise decomposition journal March 2004
Role of CD44 cytoplasmic domain in hyaluronan binding journal February 1995
Measurement of protein by spectrophotometry at 205 nm journal May 1974
A Small-Angle X-ray Scattering Apparatus for Studying Biological Macromolecules in Solution journal August 1998
CD44 Interaction with c-Src Kinase Promotes Cortactin-mediated Cytoskeleton Function and Hyaluronic Acid-dependent Ovarian Tumor Cell Migration journal November 2000
Analytical Ultracentrifugation: Sedimentation Velocity and Sedimentation Equilibrium book January 2008
Lipid Raft Association Restricts CD44-Ezrin Interaction and Promotion of Breast Cancer Cell Migration journal December 2012
Prospective Identification of Tumorigenic Prostate Cancer Stem Cells journal December 2005
Protein Identification and Analysis Tools on the ExPASy Server book January 2005
Structural Basis for CD44 Recognition by ERM Proteins journal August 2008
Identification of a phosphatidylinositol-4,5-bisphosphate-binding domain in the N-terminal region of ezrin journal December 1995
Organizing the cell cortex: the role of ERM proteins journal April 2010
Phosphoinositides in cell regulation and membrane dynamics journal October 2006
CD44 Associates With EGFR and erbB2 in Metastasizing Mammary Carcinoma Cells journal January 2002
Ezrin regulates microvillus morphogenesis by promoting distinct activities of Eps8 proteins journal March 2012
Differential roles of PtdIns(4,5)P2 and phosphorylation in moesin activation during Drosophila development journal June 2010
Neutron small-angle scattering of biological macromolecules in solution journal April 1974
CD44 exhibits a cell type dependent interaction with triton X-100 insoluble, lipid rich, plasma membrane domains journal September 1995

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Self assembly of HIV-1 Gag protein on lipid membranes generates PI(4,5)P2/Cholesterol nanoclusters posted_content October 2016
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