Conformational switching of the pseudokinase domain promotes human MLKL tetramerization and cell death by necroptosis

EJ Petrie; JJ Sandow; AV Jacobsen; BJ Smith; MDW Griffin; IS Lucet; W Dai; SN Young; MC Tanzer; A Wardak; LY Liang; AD Cowan; JM Hildebrand; WJA Kersten; G Lessene; J Silke; PE Czabotar; AI Webb; JM Murphy

Journal article

Conformational switching of the pseudokinase domain promotes human MLKL tetramerization and cell death by necroptosis

EJ Petrie, JJ Sandow, AV Jacobsen, BJ Smith, MDW Griffin, IS Lucet, W Dai, SN Young, MC Tanzer, A Wardak, LY Liang, AD Cowan, JM Hildebrand, WJA Kersten, G Lessene, J Silke, PE Czabotar, AI Webb, JM Murphy

Nature Communications | NATURE PUBLISHING GROUP | Published : 2018

DOI: 10.1038/s41467-018-04714-7

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Abstract

Necroptotic cell death is mediated by the most terminal known effector of the pathway, MLKL. Precisely how phosphorylation of the MLKL pseudokinase domain activation loop by the upstream kinase, RIPK3, induces unmasking of the N-terminal executioner four-helix bundle (4HB) domain of MLKL, higher-order assemblies, and permeabilization of plasma membranes remains poorly understood. Here, we reveal the existence of a basal monomeric MLKL conformer present in human cells prior to exposure to a necroptotic stimulus. Following activation, toggling within the MLKL pseudokinase domain promotes 4HB domain disengagement from the pseudokinase domain αC helix and pseudocatalytic loop, to enable formatio..

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University of Melbourne Researchers

Michael Griffin Author

Isabelle Lucet Author

Joanne Hildebrand Author

Guillaume Lessene Author

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Awarded by Australian Research Council

Funding Acknowledgements

We thank staff at the Australian Synchrotron SAXS/WAXS and MX beamlines for assistance with data collection, the Collaborative Crystallisation Centre (Parkville) for assistance with protein crystallization, Patrick Hennessy for technical assistance and Adeline Robin for MX data collection. We acknowledge Australian Government Research Training Program Stipend Scholarship support to A.V.J. and a Victorian International Research Scholarship to M.C.T. We are grateful to the National Health and Medical Research Council for fellowship (G.L., 1117089; J.S., 1058190; P.E.C., 1079700; J.M.M., 1105754), grant (1057905; 1124735), and infrastructure (IRIISS 9000433) support; the Australian Research Council (Future Fellowship to M.D.W.G., FT140100544); Australian Cancer Research Foundation; and the Victorian Government Operational Infrastructure Support scheme.