engleski

Structural elucidation of DISC1 pathway proteins using electron microscopy, chemical cross-linking and mass spectroscopy

Background: Disrupted-In-Schizophrenia 1 (DISC1) is a large multi- functional scaffold protein that was initially linked to schizophrenia, bipolar disorder and major depression through a rare translocation event in a Scot- tish family, but has since been associated with major mental illness in many populations worldwide. DISC1 binds several other proteins that have been independently associated with schizophrenia, including the key neurodevelopmental proteins NDE1 (Nuclear Distribution Element 1) and its paralogue NDEL1 (NDE-Like 1). A detailed understanding of the three- dimensional (3-D) structure of these p roteins would greatly facilitate the rational design of therapeutics. However, large sections of protein disorder predicted to reside in DISC1, and to a lesser extent NDE1 and NDEL1, would preclude X-ray crystallisation of these native full length proteins. Methods: Disrupted-In-Schizophrenia 1 (DISC1) is a large multi-functional scaffold protein that was initially linked to schizophrenia, bipolar disorder and major depression through a rare translocation event in a Scottish family, but has since been associated with major mental illness in many populations worldwide. DISC1 binds several other proteins that have been independently associated with schizophrenia, including the key neurodevel- opmental proteins NDE1 (Nuclear Distribution Element 1) and its paralogue NDEL1 (NDE-Like 1). A detailed understanding of the three-dimensional (3-D) structure of these proteins would greatly facilitate the rational design of therapeutics. However, large sections of protein disorder predicted to reside in DISC1, and to a lesser extent NDE1 and NDEL1, would preclude X-ray crystallisation of these native full length proteins. Results: We reveal that the native architecture of NDE1 and NDEL1 extends beyond the previous delineation of their N-terminal coiled-coil domains. We show that their partly disordered C-terminal regions facilitate a “bent- back” conformation. We also gain insight into the oligomeric state of full-length DISC1, and have uncovered the first details of the internal architecture of the protein. Discussion: The bent-backed nature of NDE1/NDEL1 reveals the importance of the C-terminus in conferring stability, sheds light on important protein- protein interactions and the pathological consequences of known clinical mutations. It also sheds light on the mechanism by which NDE1/NDEL1 interact with the key motor protein dynein. Import insights from analysis of DISC1 include the identification of novel self-association sequences and the discovery that sections of the DISC1 C-terminus, disrupted in the original Scottish family, structurally interact with the N-terminal region of the protein, with consequences for the folding of any aberrant protein derived from the translocation chromosome. 15:30 STRUCTURAL ELUCIDATION OF DISC1 PATHWAY PROTEINS USING ELECTRON MICROSCOPY, CHEMICAL CROSS-LINKING AND MASS SPECTROSCOPY. Available from: https://www.researchgate.net/publication/273423169_1530_STRUCTURAL_ELUCIDATION_OF_DISC1_PATHWAY_PROTEINS_USING_ELECTRON_MICROSCOPY_CHEMICAL_CROSS-LINKING_AND_MASS_SPECTROSCOPY [accessed Jul 20, 2017].

DISC1 ; NDE1 ; structural biology ; biophysics ; schizophrenia

nije evidentirano