Biophysical insights from a single chain camelid antibody directed against the Disrupted-in-Schizophrenia 1 protein

PLoS One. 2018 Jan 11;13(1):e0191162. doi: 10.1371/journal.pone.0191162. eCollection 2018.

Abstract

Accumulating evidence suggests an important role for the Disrupted-in-Schizophrenia 1 (DISC1) protein in neurodevelopment and chronic mental illness. In particular, the C-terminal 300 amino acids of DISC1 have been found to mediate important protein-protein interactions and to harbor functionally important phosphorylation sites and disease-associated polymorphisms. However, long disordered regions and oligomer-forming subdomains have so far impeded structural analysis. VHH domains derived from camelid heavy chain only antibodies are minimal antigen binding modules with appreciable solubility and stability, which makes them well suited for the stabilizing proteins prior to structural investigation. Here, we report on the generation of a VHH domain derived from an immunized Lama glama, displaying high affinity for the human DISC1 C region (aa 691-836), and its characterization by surface plasmon resonance, size exclusion chromatography and immunological techniques. The VHH-DISC1 (C region) complex was also used for structural investigation by small angle X-ray scattering analysis. In combination with molecular modeling, these data support predictions regarding the three-dimensional fold of this DISC1 segment as well as its steric arrangement in complex with our VHH antibody.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antigen-Antibody Complex / chemistry
  • Antigen-Antibody Complex / genetics
  • Antigen-Antibody Reactions
  • Biophysical Phenomena
  • Camelids, New World / genetics
  • Camelids, New World / immunology*
  • Epitope Mapping
  • Female
  • Humans
  • Immunoglobulin Heavy Chains / chemistry
  • Immunoglobulin Heavy Chains / genetics
  • Mice
  • Models, Molecular
  • Nerve Tissue Proteins / chemistry
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / immunology*
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Fragments / immunology
  • Protein Interaction Domains and Motifs
  • Scattering, Small Angle
  • Single-Chain Antibodies / chemistry*
  • Single-Chain Antibodies / genetics
  • Surface Plasmon Resonance
  • X-Ray Diffraction

Substances

  • Antigen-Antibody Complex
  • DISC1 protein, human
  • Disc1 protein, mouse
  • Immunoglobulin Heavy Chains
  • Nerve Tissue Proteins
  • Peptide Fragments
  • Single-Chain Antibodies

Grant support

This work was supported by NRW Graduate School BioStruct (http://www.biostruct.uni-duesseldorf.de/startseite-biostruct.html) to CK; HHU Graduate School iBRAIN (http://www.ibrain-duesseldorf.de) to CK; Fritz-Thyssen Foundation (http://www.fritz-thyssen-stiftung.de/?L=1) (#10.14.2.140) to NJB; and EU-FP7 MC-ITN IN-SENS (#60761) to CK.