title: The BECN1 N-terminal domain is intrinsically disordered.
abstract: BECN1/Beclin 1 has a critical role in the early stages of autophagosome formation. Recently, structures of its central and C-terminal domains were reported, however, little structural information is available on the N-terminal domain, comprising a third of the protein. This lack of structural information largely stems from the inability to produce this region in a purified form. Here, we describe the expression and purification of the N-terminal domain of BECN1 (residues 1 to 150) and detailed biophysical characterization, including NMR spectroscopy. Combined, our studies demonstrated at the atomic level that the BECN1 N-terminal domain is intrinsically disordered, and apart from the BH3 subdomain, remains disordered following interaction with a binding partner, BCL2L1/BCL-XL. In addition, the BH3 domain α-helix induced upon interaction with BCL2L1 reverts to a disordered state when the complex is dissociated by exposure to a competitive inhibitor. No significant interactions between N- and C-terminal domains were detected.
authors.affiliation: a The Walter and Eliza Hall Institute of Medical Research , Parkville , Victoria , Australia.|b Department of Medical Biology , The University of Melbourne , Parkville , Victoria , Australia.|c Olivia Newton-John Cancer Research Institute , Heidelberg , Victoria , Australia.|d School of Cancer Medicine, La Trobe University , Melbourne , Victoria , Australia.|e Department of Chemistry and Physics , La Trobe Institute for Molecular Science , Melbourne , Victoria , Australia.
authors.forename: Erinna F
authors.initials: EF
authors.lastname: Lee
authors.affiliation: f Department of Biochemistry and Genetics , La Trobe Institute for Molecular Science, La Trobe University , Melbourne , Victoria , Australia.
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Perugini
authors.affiliation: a The Walter and Eliza Hall Institute of Medical Research , Parkville , Victoria , Australia.
authors.forename: Anne
authors.initials: A
authors.lastname: Pettikiriarachchi
authors.affiliation: a The Walter and Eliza Hall Institute of Medical Research , Parkville , Victoria , Australia.|c Olivia Newton-John Cancer Research Institute , Heidelberg , Victoria , Australia.
authors.forename: Marco
authors.initials: M
authors.lastname: Evangelista
authors.affiliation: g Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne , Parkville , Victoria , Australia.
authors.forename: David W
authors.initials: DW
authors.lastname: Keizer
authors.affiliation: g Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne , Parkville , Victoria , Australia.
authors.forename: Shenggen
authors.initials: S
authors.lastname: Yao
authors.affiliation: a The Walter and Eliza Hall Institute of Medical Research , Parkville , Victoria , Australia.|b Department of Medical Biology , The University of Melbourne , Parkville , Victoria , Australia.|c Olivia Newton-John Cancer Research Institute , Heidelberg , Victoria , Australia.|d School of Cancer Medicine, La Trobe University , Melbourne , Victoria , Australia.|e Department of Chemistry and Physics , La Trobe Institute for Molecular Science , Melbourne , Victoria , Australia.
authors.forename: W Douglas
authors.initials: WD
authors.lastname: Fairlie
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D017209:Apoptosis; D000071186:Beclin-1 / Q000737:chemistry*; D002942:Circular Dichroism; D006801:Humans; D064267:Intrinsically Disordered Proteins / Q000737:chemistry*; D051379:Mice; D010766:Phosphorylation; D000072417:Protein Domains; D017433:Protein Structure, Secondary; D051020:bcl-X Protein / Q000737:chemistry
chemical_list: D000596:Amino Acids; C491997:BECN1 protein, human; D000071186:Beclin-1; D064267:Intrinsically Disordered Proteins; D051020:bcl-X Protein
keywords: BCL2; BECN1; BH3 domain; Beclin 1; autophagy; intrinsically disordered protein; nuclear magnetic resonance