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

• Published in: PloS one 2018-01, Vol.13 (1), p.e0191162-e0191162
• Main Authors: Yerabham, Antony S K, Müller-Schiffmann, Andreas, Ziehm, Tamar, Stadler, Andreas, Köber, Sabrina, Indurkhya, Xela, Marreiros, Rita, Trossbach, Svenja V, Bradshaw, Nicholas J, Prikulis, Ingrid, Willbold, Dieter, Weiergräber, Oliver H, Korth, Carsten
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Amino acids; Animals; Antibodies; Antigen-Antibody Complex - chemistry; Antigen-Antibody Complex - genetics; Antigen-Antibody Reactions; Antigens; Biochemistry; Biology and Life Sciences; Biophysical Phenomena; Camelids, New World - genetics; Camelids, New World - immunology; Chains; Chromatography; Crystallography; Development and progression; Disc1 protein; Epitope Mapping; Female; Genetic aspects; Humans; Immunization; Immunoglobulin Heavy Chains - chemistry; Immunoglobulin Heavy Chains - genetics; Immunoglobulins; Immunology; Medicine and Health Sciences; Mental disorders; Mice; Models, Molecular; Molecular biology; Molecular modelling; Nerve tissue proteins; Nerve Tissue Proteins - chemistry; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - immunology; Neuropathology; Neurosciences; Peptide Fragments - chemistry; Peptide Fragments - genetics; Peptide Fragments - immunology; Phosphorylation; Physical Sciences; Physiological aspects; Protein interaction; Protein Interaction Domains and Motifs; Protein-protein interactions; Proteins; Research and Analysis Methods; Scattering, Small Angle; Schizophrenia; Single-Chain Antibodies - chemistry; Single-Chain Antibodies - genetics; Size exclusion chromatography; Small angle X ray scattering; Solubility; Structural analysis; Surface Plasmon Resonance; Three dimensional models; X ray scattering; X-Ray Diffraction
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0191162

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.