Family-wide Structural and Biophysical Analysis of Binding Interactions among Non-clustered δ-Protocadherins
Oliver J. Harrison,
Julia Brasch,
Phinikoula S. Katsamba,
Goran Ahlsen,
Alex J. Noble,
Hanbin Dan,
Rosemary V. Sampogna,
Clinton S. Potter,
Bridget Carragher,
Barry Honig,
Lawrence Shapiro
Affiliations
Oliver J. Harrison
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032, USA; Zuckerman Mind Brain Behavior Institute, Columbia University, New York, NY 10027, USA
Julia Brasch
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032, USA; Zuckerman Mind Brain Behavior Institute, Columbia University, New York, NY 10027, USA; National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, NY 10027, USA
Phinikoula S. Katsamba
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032, USA; Zuckerman Mind Brain Behavior Institute, Columbia University, New York, NY 10027, USA
Goran Ahlsen
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032, USA; Zuckerman Mind Brain Behavior Institute, Columbia University, New York, NY 10027, USA
Alex J. Noble
National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, NY 10027, USA
Hanbin Dan
Department of Medicine, Division of Nephrology, Columbia University, New York, NY 10032, USA
Rosemary V. Sampogna
Department of Medicine, Division of Nephrology, Columbia University, New York, NY 10032, USA
Clinton S. Potter
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032, USA; National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, NY 10027, USA
Bridget Carragher
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032, USA; National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, NY 10027, USA
Barry Honig
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032, USA; Zuckerman Mind Brain Behavior Institute, Columbia University, New York, NY 10027, USA; Department of Medicine, Division of Nephrology, Columbia University, New York, NY 10032, USA; Department of Systems Biology, Columbia University, New York, NY 10032, USA; Corresponding author
Lawrence Shapiro
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032, USA; Zuckerman Mind Brain Behavior Institute, Columbia University, New York, NY 10027, USA; Corresponding author
Summary: Non-clustered δ1- and δ2-protocadherins, close relatives of clustered protocadherins, function in cell adhesion and motility and play essential roles in neural patterning. To understand the molecular interactions underlying these functions, we used solution biophysics to characterize binding of δ1- and δ2-protocadherins, determined crystal structures of ectodomain complexes from each family, and assessed ectodomain assembly in reconstituted intermembrane junctions by cryoelectron tomography (cryo-ET). Homophilic trans (cell–cell) interactions were preferred for all δ-protocadherins, with additional weaker heterophilic interactions observed exclusively within each subfamily. As expected, δ1- and δ2-protocadherin trans dimers formed through antiparallel EC1–EC4 interfaces, like clustered protocadherins. However, no ectodomain-mediated cis (same-cell) interactions were detectable in solution; consistent with this, cryo-ET of reconstituted junctions revealed dense assemblies lacking the characteristic order observed for clustered protocadherins. Our results define non-clustered protocadherin binding properties and their structural basis, providing a foundation for interpreting their functional roles in neural patterning. : Non-clustered δ-protocadherins are adhesion molecules linked to a number of neurological disorders. Harrison et al. apply biophysical and structural methods across the family to show preferential self-binding through a canonical interface and disordered assemblies of adhesive dimers between membranes that diverge from ordered assemblies of close relatives clustered protocadherins. Keywords: cell adhesion, non-clustered protocadherins, cadherin, X-ray crystallography, cryo-electron tomography, homophilic adhesion, neuronal cell adhesion
