title: Structural insights into the activation mechanism of dynamin-like EHD ATPases.
abstract: Eps15 (epidermal growth factor receptor pathway substrate 15)-homology domain containing proteins (EHDs) comprise a family of dynamin-related mechano-chemical ATPases involved in cellular membrane trafficking. Previous studies have revealed the structure of the EHD2 dimer, but the molecular mechanisms of membrane recruitment and assembly have remained obscure. Here, we determined the crystal structure of an amino-terminally truncated EHD4 dimer. Compared with the EHD2 structure, the helical domains are 50° rotated relative to the GTPase domain. Using electron paramagnetic spin resonance (EPR), we show that this rotation aligns the two membrane-binding regions in the helical domain toward the lipid bilayer, allowing membrane interaction. A loop rearrangement in GTPase domain creates a new interface for oligomer formation. Our results suggest that the EHD4 structure represents the active EHD conformation, whereas the EHD2 structure is autoinhibited, and reveal a complex series of domain rearrangements accompanying activation. A comparison with other peripheral membrane proteins elucidates common and specific features of this activation mechanism.
authors.affiliation: Crystallography Department, Max-Delbrück-Centrum for Molecular Medicine, 13125 Berlin, Germany.|Institute of Chemistry and Biochemistry, Freie Universität Berlin, 14195 Berlin, Germany.
authors.forename: Arthur Alves
authors.initials: AA
authors.lastname: Melo
authors.affiliation: Department of Physics, Rani Channamma University, 591 156, Karnataka, India.
authors.forename: Balachandra G
authors.initials: BG
authors.lastname: Hegde
authors.affiliation: Crystallography Department, Max-Delbrück-Centrum for Molecular Medicine, 13125 Berlin, Germany.|Institute of Chemistry and Biochemistry, Freie Universität Berlin, 14195 Berlin, Germany.
authors.forename: Claudio
authors.initials: C
authors.lastname: Shah
authors.affiliation: Medical Biochemistry and Biophysics, Umeå University, 901 87 Umeå, Sweden.
authors.forename: Elin
authors.initials: E
authors.lastname: Larsson
authors.affiliation: Zilkha Neurogenetic Institute, University of Southern California, Los Angeles, CA 90033.
authors.forename: J Mario
authors.initials: JM
authors.lastname: Isas
authors.affiliation: Crystallography Department, Max-Delbrück-Centrum for Molecular Medicine, 13125 Berlin, Germany.
authors.forename: Séverine
authors.initials: S
authors.lastname: Kunz
authors.affiliation: Medical Biochemistry and Biophysics, Umeå University, 901 87 Umeå, Sweden.|Integrative Medical Biology, Umeå University, 901 87, Umeå, Sweden.
authors.forename: Richard
authors.initials: R
authors.lastname: Lundmark
authors.affiliation: Zilkha Neurogenetic Institute, University of Southern California, Los Angeles, CA 90033.
authors.forename: Ralf
authors.initials: R
authors.lastname: Langen
authors.affiliation: Crystallography Department, Max-Delbrück-Centrum for Molecular Medicine, 13125 Berlin, Germany; oliver.daumke@mdc-berlin.de.|Institute of Chemistry and Biochemistry, Freie Universität Berlin, 14195 Berlin, Germany.
authors.forename: Oliver
authors.initials: O
authors.lastname: Daumke
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000595:Amino Acid Sequence / Q000235:genetics; D002352:Carrier Proteins / Q000378:metabolism*; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism*; D018360:Crystallography, X-Ray; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004789:Enzyme Activation / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D009687:Nuclear Proteins / Q000378:metabolism*; D011485:Protein Binding; D000072417:Protein Domains / Q000502:physiology; D055503:Protein Multimerization; D021381:Protein Transport / Q000502:physiology
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; D004268:DNA-Binding Proteins; C406446:EHD2 protein, human; C406448:EHD4 protein, human; C486206:EPS15 protein, human; D009687:Nuclear Proteins; D000251:Adenosine Triphosphatases
keywords: autoinhibition; dynamin superfamily; endocytic pathways; membrane remodeling; protein structure