PUBLICATION

Structure of protein O-mannose kinase reveals a unique active site architecture

Authors
Zhu, Q., Venzke, D., Walimbe, A.S., Anderson, M.E., Fu, Q., Kinch, L.N., Wang, W., Chen, X., Grishin, N.V., Huang, N., Yu, L., Dixon, J.E., Campbell, K.P., Xiao, J.
ID
ZDB-PUB-161124-9
Date
2016
Source
eLIFE   5: (Journal)
Registered Authors
Keywords
biochemistry, biophysics, none, structural biology
MeSH Terms
  • Protein Interaction Domains and Motifs
  • Cloning, Molecular
  • Protein Binding
  • Magnesium/chemistry*
  • Magnesium/metabolism
  • Animals
  • Adenosine Diphosphate/chemistry*
  • Adenosine Diphosphate/metabolism
  • Amino Acid Sequence
  • Crystallography, X-Ray
  • Catalytic Domain
  • Aluminum Compounds/chemistry
  • Protein Conformation, beta-Strand
  • Trisaccharides/chemistry*
  • Trisaccharides/metabolism
  • Recombinant Proteins/chemistry
  • Recombinant Proteins/genetics
  • Recombinant Proteins/metabolism
  • Sequence Homology, Amino Acid
  • Fluorides/chemistry
  • Protein Kinases/chemistry*
  • Protein Kinases/genetics
  • Protein Kinases/metabolism
  • Baculoviridae/genetics
  • Baculoviridae/metabolism
  • Mutation
  • Mannose/chemistry*
  • Mannose/metabolism
  • Humans
  • Sf9 Cells
  • Gene Expression
  • Dystroglycans/chemistry*
  • Dystroglycans/metabolism
  • Protein Conformation, alpha-Helical
  • Fish Proteins/chemistry*
  • Fish Proteins/genetics
  • Fish Proteins/metabolism
  • Sequence Alignment
  • Zebrafish/metabolism
  • Substrate Specificity
  • Models, Molecular
  • Phosphorylation
(all 42)
PubMed
27879205 Full text @ Elife
Abstract
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.
Genes / Markers
Figures
No images available
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Your Input Welcome
We welcome your input and comments. Please use this form to recommend updates to the information in ZFIN. We appreciate as much detail as possible and references as appropriate. We will review your comments promptly.
Citation
Zhu, Q., Venzke, D., Walimbe, A.S., Anderson, M.E., Fu, Q., Kinch, L.N., Wang, W., Chen, X., Grishin, N.V., Huang, N., Yu, L., Dixon, J.E., Campbell, K.P., Xiao, J. (2016) Structure of protein O-mannose kinase reveals a unique active site architecture. eLIFE. 5.