Structure of protein O-mannose kinase reveals a unique active site architecture
Copyright policy )Structure of protein O-mannose kinase reveals a unique active site architecture
The ‘pseudokinase’ SgK196 is a protein O-mannose kinase (POMK) that catalyzes an essential phosphorylation step during biosynthesis of the laminin-binding glycan on α-dystroglycan. However, the catalytic mechanism underlying this activity remains elusive. Here we present the crystal structure of Danio rerio POMK in complex with Mg2+ ions, ADP, aluminum fluoride, and the GalNAc-β3-GlcNAc-β4-Man trisaccharide substrate, thereby providing a snapshot of the catalytic transition state of this unusual kinase. The active site of POMK is established by residues located in non-canonical positions and is stabilized by a disulfide bridge. GalNAc-β3-GlcNAc-β4-Man is recognized by a surface groove, and the GalNAc-β3-GlcNAc moiety mediates the majority of interactions with POMK. Expression of various POMK mutants in POMK knockout cells further validated the functional requirements of critical residues. Our results provide important insights into the ability of POMK to function specifically as a glycan kinase, and highlight the structural diversity of the human kinome.
- Tsinghua University China (People's Republic of)
- University of California, San Diego United States
- Peking University China (People's Republic of)
- University of Iowa United States
- National Institute of Biological Sciences, Beijing China (People's Republic of)
Models, Molecular, Protein Conformation, alpha-Helical, Biomedical and clinical sciences, Protein Conformation, none, Sequence Homology, Gene Expression, Crystallography, X-Ray, Biochemistry, Substrate Specificity, Fluorides, Models, biophysics, Catalytic Domain, Sf9 Cells, structural biology, Magnesium, Biology (General), Cloning, Molecular, Phosphorylation, Aluminum Compounds, Dystroglycans, Zebrafish, Crystallography, secretory pathway kinase, Q, R, dystroglycan biosynthesis, Biological Sciences, Recombinant Proteins, Adenosine Diphosphate, Amino Acid, Biological sciences, Medicine, Baculoviridae, Protein Binding, muscular dystrophy, Fish Proteins, QH301-705.5, 1.1 Normal biological development and functioning, Science, Underpinning research, biochemistry, Animals, Humans, Protein Interaction Domains and Motifs, Amino Acid Sequence, alpha-Helical, Molecular, Health sciences, Mutation, X-Ray, beta-Strand, Biochemistry and Cell Biology, Generic health relevance, Protein Kinases, Sequence Alignment, Trisaccharides, Mannose, Cloning
Models, Molecular, Protein Conformation, alpha-Helical, Biomedical and clinical sciences, Protein Conformation, none, Sequence Homology, Gene Expression, Crystallography, X-Ray, Biochemistry, Substrate Specificity, Fluorides, Models, biophysics, Catalytic Domain, Sf9 Cells, structural biology, Magnesium, Biology (General), Cloning, Molecular, Phosphorylation, Aluminum Compounds, Dystroglycans, Zebrafish, Crystallography, secretory pathway kinase, Q, R, dystroglycan biosynthesis, Biological Sciences, Recombinant Proteins, Adenosine Diphosphate, Amino Acid, Biological sciences, Medicine, Baculoviridae, Protein Binding, muscular dystrophy, Fish Proteins, QH301-705.5, 1.1 Normal biological development and functioning, Science, Underpinning research, biochemistry, Animals, Humans, Protein Interaction Domains and Motifs, Amino Acid Sequence, alpha-Helical, Molecular, Health sciences, Mutation, X-Ray, beta-Strand, Biochemistry and Cell Biology, Generic health relevance, Protein Kinases, Sequence Alignment, Trisaccharides, Mannose, Cloning
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