Characterisation of the conformational preference and dynamics of the intrinsically disordered N-terminal region of Beclin 1 by NMR spectroscopy.
Beclin 1 is a 450 amino acid protein that plays critical roles in the early stages of autophagosome formation. We recently reported the successful expression, purification and structural characterisation of the entire N-terminal region of Beclin 1 (residues 1-150), including its backbone NMR chemical shift assignments. Based on assigned backbone NMR chemical shifts, it has been established that the N-terminal region of Beclin 1 (1-150), including the BH3 domain (112-123), is intrinsically disordered in the absence of its interaction partners. Here, a detailed study of its conformational preference and backbone dynamics obtained from an analysis of its secondary structure populations using the δ2D method, and the measurements of effective hydrodynamic radius as well as (1)H temperature coefficients, (1)H solvent exchange rates, and (15)N relaxation parameters of backbone amides using NMR spectroscopy is reported. These data provide further evidence for the intrinsically disordered nature of the N-terminal region of Beclin 1 and support the view that the helical conformation adopted by the Beclin 1 BH3 domain upon interaction with binding partners such as BCL-2 pro-survival proteins is likely induced rather than pre-existing.
Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Victoria 3010, Australia. Electronic address: shyao@unimelb.edu.au.
Shenggen
S
Yao
The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia; Department of Medical Biology, The University of Melbourne, Victoria 3010, Australia; Olivia Newton-John Cancer Research Institute, Heidelberg, Victoria 3084, Australia; School of Cancer Medicine, La Trobe University, Melbourne, Victoria 3086, Australia; Department of Chemistry and Physics, La Trobe Institute for Molecular Science, Melbourne, Victoria 3086, Australia. Electronic address: Erinna.Lee@latrobe.edu.au.
Erinna F
EF
Lee
The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia.
Anne
A
Pettikiriarachchi
The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia; Olivia Newton-John Cancer Research Institute, Heidelberg, Victoria 3084, Australia; School of Cancer Medicine, La Trobe University, Melbourne, Victoria 3086, Australia.
Marco
M
Evangelista
Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Victoria 3010, Australia.
David W
DW
Keizer
The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia; Department of Medical Biology, The University of Melbourne, Victoria 3010, Australia; Olivia Newton-John Cancer Research Institute, Heidelberg, Victoria 3084, Australia; School of Cancer Medicine, La Trobe University, Melbourne, Victoria 3086, Australia; Department of Chemistry and Physics, La Trobe Institute for Molecular Science, Melbourne, Victoria 3086, Australia.
W Douglas
WD
Fairlie
D000071186:Beclin-1 / Q000737:chemistry*; D006801:Humans; D064267:Intrinsically Disordered Proteins / Q000737:chemistry*; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D009587:Nitrogen Isotopes; D000072756:Protein Conformation, alpha-Helical; D054730:Protein Interaction Domains and Motifs; D011994:Recombinant Proteins / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods; D013816:Thermodynamics
C491997:BECN1 protein, human; D000071186:Beclin-1; D064267:Intrinsically Disordered Proteins; D009587:Nitrogen Isotopes; D011994:Recombinant Proteins
BH3 domain; Beclin 1; Conformational dynamics; Intrinsically disordered protein; NMR