6UX8
Structure of monobody 33 MLKL N-terminal domain complex
Summary for 6UX8
| Entry DOI | 10.2210/pdb6ux8/pdb |
| Descriptor | Mixed lineage kinase domain-like protein, Monobody, ZINC ION, ... (4 entities in total) |
| Functional Keywords | protein interactions, cell death, ripk3, programmed necrosis, protein engineering, transferase |
| Biological source | Homo sapiens (Human) More |
| Total number of polymer chains | 2 |
| Total formula weight | 28977.14 |
| Authors | Birkinshaw, R.W.,Petrie, E.J.,Murphy, J.M. (deposition date: 2019-11-07, release date: 2020-04-01, Last modification date: 2023-10-11) |
| Primary citation | Petrie, E.J.,Birkinshaw, R.W.,Koide, A.,Denbaum, E.,Hildebrand, J.M.,Garnish, S.E.,Davies, K.A.,Sandow, J.J.,Samson, A.L.,Gavin, X.,Fitzgibbon, C.,Young, S.N.,Hennessy, P.J.,Smith, P.P.C.,Webb, A.I.,Czabotar, P.E.,Koide, S.,Murphy, J.M. Identification of MLKL membrane translocation as a checkpoint in necroptotic cell death using Monobodies. Proc.Natl.Acad.Sci.USA, 117:8468-8475, 2020 Cited by PubMed Abstract: The necroptosis cell death pathway has been implicated in host defense and in the pathology of inflammatory diseases. While phosphorylation of the necroptotic effector pseudokinase Mixed Lineage Kinase Domain-Like (MLKL) by the upstream protein kinase RIPK3 is a hallmark of pathway activation, the precise checkpoints in necroptosis signaling are still unclear. Here we have developed monobodies, synthetic binding proteins, that bind the N-terminal four-helix bundle (4HB) "killer" domain and neighboring first brace helix of human MLKL with nanomolar affinity. When expressed as genetically encoded reagents in cells, these monobodies potently block necroptotic cell death. However, they did not prevent MLKL recruitment to the "necrosome" and phosphorylation by RIPK3, nor the assembly of MLKL into oligomers, but did block MLKL translocation to membranes where activated MLKL normally disrupts membranes to kill cells. An X-ray crystal structure revealed a monobody-binding site centered on the α4 helix of the MLKL 4HB domain, which mutational analyses showed was crucial for reconstitution of necroptosis signaling. These data implicate the α4 helix of its 4HB domain as a crucial site for recruitment of adaptor proteins that mediate membrane translocation, distinct from known phospholipid binding sites. PubMed: 32234780DOI: 10.1073/pnas.1919960117 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.5 Å) |
Structure validation
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