crystal structure of the n-acetylmannosamine kinase domain of gne带有n-acetylmannosamine激酶结构域的晶体结构.pdfVIP

Crystal Structure of the N-Acetylmannosamine Kinase Domain of GNE 1 1 1 1 1,2 Yufeng Tong , Wolfram Tempel , Lyudmila Nedyalkova , Farrell MacKenzie , Hee-Won Park * 1 Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada, 2 Department of Pharmacology, University of Toronto, Toronto, Ontario, Canada Abstract Background: UDP-GlcNAc 2-epimerase/ManNAc 6-kinase, GNE, is a bi-functional enzyme that plays a key role in sialic acid biosynthesis. Mutations of the GNE protein cause sialurea or autosomal recessive inclusion body myopathy/Nonaka myopathy. GNE is the only human protein that contains a kinase domain belonging to the ROK (repressor, ORF, kinase) family. Principal Findings: We solved the structure of the GNE kinase domain in the ligand-free state. The protein exists predominantly as a dimer in solution, with small populations of monomer and higher-order oligomer in equilibrium with the dimer. Crystal packing analysis reveals the existence of a crystallographic hexamer, and that the kinase domain dimerizes through the C-lobe subdomain. Mapping of disease-related missense mutations onto the kinase domain structure revealed that the mutation sites could be classified into four different groups based on the location – dimer interface, interlobar helices, protein surface, or within other secondary structural elements. Conclusions: The crystal structure of the kinase domain of GNE provides a structural basis for understanding