4a5a | pdb_00004a5a

Crystal structure of the C258S/C268S variant of Toxoplasma gondii nucleoside triphosphate diphosphohydrolase 3 (NTPDase3) in complex with magnesium and AMPPNP

spinqualitylabelsall models 4a5a, resolution 2.85Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 4a5a is a 4 chain structure with sequence from Toxgo. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Ligands: ANP, MG Related: 4a57, 4a59, 4a5b Activity: Apyrase, with EC number 3.6.1.5 Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function [NTP1_TOXGO] May perform an important processing step in the conversion of high energy nucleotides prior to uptake by the parasite and may contribute to intracellular survival and virulence. NTPAse-I has a specific activity 4.5-fold higher than NTPAse-II in hydrolysis of ATP. The primary difference between these isozymes lies in their ability to hydrolyze nucleoside triphosphate versus diphosphate substrates. While NTPAse-II hydrolyzes ATP to ADP and ADP to AMP at almost the same rate, NTPAse-I hydrolyzes ADP to AMP at a much slower rate (0.7% of the rate for ATP). Publication Abstract from PubMed The intracellular parasite Toxoplasma gondii produces two nucleoside triphosphate diphosphohydrolases (NTPDase1 and -3). These tetrameric, cysteine-rich enzymes require activation by reductive cleavage of a hitherto unknown disulfide bond. Despite a 97% sequence identity both isozymes differ largely in their ability to hydrolyze ATP and ADP. Here, we present crystal structures of inactive NTPDase3 as apo form and in complex with the product AMP to resolutions of 2.0 Ang. and 2.2 Ang., respectively. We find that the enzyme is present in an open conformation that precludes productive substrate binding and catalysis. The cysteine bridge 258-268 is identified to be responsible for locking of activity. Crystal structures of constitutively active variants of NTPDase1 and -3 generated by mutation of C258-C268 show that opening of the regulatory cysteine bridge induces a pronounced contraction of the whole tetramer. This is accompanied by a 12 deg domain closure motion resulting in the correct arrangement of all active site residues. A complex structure of activated NTPDase3 with a non-hydrolyzable ATP analog and the cofactor Mg2+ to a resolution of 2.85 Ang. indicates that catalytic differences between the NTPDases are primarily dictated by differences in positioning of the adenine base caused by substitution of R492 and E493 in NTPDase1 by glycines in NTPDase3. Structural insight into the activation mechanism of Toxoplasma gondii nucleoside triphosphate diphosphohydrolases by disulfide reduction.,Krug U, Zebisch M, Krauss M, Strater N J Biol Chem. 2011 Nov 30. PMID:22130673 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See Also Ectonucleoside triphosphate diphosphohydrolase Nucleoside triphosphatase Sandbox 30008

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