Insights into the evolution of divergent nucleotide-binding mechanisms among pseudokinases revealed by crystal structures of human and mouse MLKL.
The pseudokinase MLKL (mixed lineage kinase domain-like) was identified recently as an essential checkpoint in the programmed necrosis or 'necroptosis' cell death pathway. In the present study, we report the crystal structure of the human MLKL pseudokinase domain at 1.7 Å (1 Å=0.1 nm) resolution and probe its nucleotide-binding mechanism by performing structure-based mutagenesis. By comparing the structures and nucleotide-binding determinants of human and mouse MLKL orthologues, the present study provides insights into the evolution of nucleotide-binding mechanisms among pseudokinases and their mechanistic divergence from conventional catalytically active protein kinases.
James M
JM
Murphy
‡Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Clayton, Victoria 3800, Australia.
Isabelle S
IS
Lucet
Joanne M
JM
Hildebrand
Maria C
MC
Tanzer
*The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia.
Samuel N
SN
Young
Pooja
P
Sharma
Guillaume
G
Lessene
Warren S
WS
Alexander
Jeffrey J
JJ
Babon
John
J
Silke
Peter E
PE
Czabotar
D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism*; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D001665:Binding Sites; D018360:Crystallography, X-Ray; D030562:Databases, Protein; D019143:Evolution, Molecular; D006801:Humans; D008239:Lysine / Q000737:chemistry; D051379:Mice; D008958:Models, Molecular; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism; D015202:Protein Engineering; D011494:Protein Kinases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D055550:Protein Stability; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D053838:Scattering, Small Angle; D016415:Sequence Alignment
D050505:Mutant Proteins; D011994:Recombinant Proteins; D000255:Adenosine Triphosphate; C570693:MLKL protein, human; C570694:MLKL protein, mouse; D011494:Protein Kinases; D008239:Lysine