title: Crystal structure of a soluble form of human CD73 with ecto-5'-nucleotidase activity.
abstract: CD73 is a dimeric ecto-5'-nucleotidase that is expressed on the exterior side of the plasma membrane. CD73 has important regulatory functions in the extracellular metabolism of certain nucleoside monophosphates, in particular adenosine monophosphate, and has been linked to a number of pathological conditions such as cancer and myocardial ischaemia. Here, we present the crystal structure of a soluble form of human soluble CD73 (sCD73) at 2.2 Å resolution, a truncated form of CD73 that retains ecto-5'-nucleotidase activity. With this structure we obtained insight into the dimerisation of CD73, active site architecture, and a sense of secondary modifications of the protein. The crystal structure reveals a conserved loop that is directly involved in the dimer-dimer interaction showing that the two subunits of the dimer are not linked by disulfide bridges. Using biophotonic microarray imaging we were able to confirm glycosylation of the enzyme and show that the enzyme is decorated with a variety of oligosaccharide structures. The crystal structure of sCD73 will aid the design of inhibitors or activator molecules for the treatment of several diseases and prove useful in explaining the possible roles of single nucleotide polymorphisms in physiology and disease.
authors.affiliation: Manchester Institute of Biotechnology, University of Manchester, 131 Princess Street, Manchester M1 7DN, UK.
authors.forename: Dominic P H M
authors.initials: DP
authors.lastname: Heuts
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Weissenborn
authors.forename: Rouslan V
authors.initials: RV
authors.lastname: Olkhov
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Shaw
authors.forename: Jennet
authors.initials: J
authors.lastname: Gummadova
authors.forename: Colin
authors.initials: C
authors.lastname: Levy
authors.forename: Nigel S
authors.initials: NS
authors.lastname: Scrutton
mesh_terms: D015720:5'-Nucleotidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000201:enzymology; D058851:GPI-Linked Proteins / Q000737:chemistry; D006801:Humans; D008958:Models, Molecular; D016415:Sequence Alignment; D012995:Solubility
chemical_list: D058851:GPI-Linked Proteins; D015720:5'-Nucleotidase; C548241:NT5E protein, human