3FOW

Plasmodium Purine Nucleoside Phosphorylase V66I-V73I-Y160F Mutant

Summary for 3FOW

• Entry DOI: 10.2210/pdb3fow/pdb
• Related: 1NW4
• Descriptor: Uridine phosphorylase, putative, PHOSPHATE ION, 1,4-DIDEOXY-4-AZA-1-(S)-(9-DEAZAHYPOXANTHIN-9-YL)-D-RIBITOL, ... (4 entities in total)
• Functional Keywords: protein-inhibitor complex, phosphorylase, glycosyltransferase, transferase
• Biological source: Plasmodium falciparum
• Total number of polymer chains: 2
• Total formula weight: 61940.58

Authors

Donaldson, T.,Zhan, C. (deposition date: 2009-01-02, release date: 2009-12-29, Last modification date: 2023-09-20)

Primary citation

Donaldson, T.M.,Ting, L.M.,Zhan, C.,Shi, W.,Zheng, R.,Almo, S.C.,Kim, K.
Structural determinants of the 5'-methylthioinosine specificity of Plasmodium purine nucleoside phosphorylase.
Plos One, 9:e84384-e84384, 2014

PubMed Abstract: Plasmodium parasites rely upon purine salvage for survival. Plasmodium purine nucleoside phosphorylase is part of the streamlined Plasmodium purine salvage pathway that leads to the phosphorylysis of both purines and 5'-methylthiopurines, byproducts of polyamine synthesis. We have explored structural features in Plasmodium falciparum purine nucleoside phosphorylase (PfPNP) that affect efficiency of catalysis as well as those that make it suitable for dual specificity. We used site directed mutagenesis to identify residues critical for PfPNP catalytic activity as well as critical residues within a hydrophobic pocket required for accommodation of the 5'-methylthio group. Kinetic analysis data shows that several mutants had disrupted binding of the 5'-methylthio group while retaining activity for inosine. A triple PfPNP mutant that mimics Toxoplasma gondii PNP had significant loss of 5'-methylthio activity with retention of inosine activity. Crystallographic investigation of the triple mutant PfPNP with Tyr160Phe, Val66Ile, andVal73Ile in complex with the transition state inhibitor immucillin H reveals fewer hydrogen bond interactions for the inhibitor in the hydrophobic pocket.

PubMed: 24416224
DOI: 10.1371/journal.pone.0084384

Experimental method

X-RAY DIFFRACTION (2.8 Å)