Purine nucleoside phosphorylases: properties, functions, and clinical aspects.
The ubiquitous purine nucleoside phosphorylases (PNPs) play a key role in the purine salvage pathway, and PNP deficiency in humans leads to an impairment of T-cell function, usually with no apparent effects on B-cell function. This review updates the properties of the enzymes from eukaryotes and a wide range of prokaryotes, including a tentative classification of the enzymes from various sources, based on three-dimensional structures in the solid state, subunit composition, amino acid sequences, and substrate specificities. Attention is drawn to the compelling need of quantitative experimental data on subunit composition in solution, binding constants, and stoichiometry of binding; order of ligand binding and release; and its possible relevance to the complex kinetics exhibited with some substrates. Mutations responsible for PNP deficiency are described, as well as clinical methods, including gene therapy, for corrections of this usually fatal disease. Substrate discrimination between enzymes from different sources is also being profited from for development of tumour-directed gene therapy. Detailed accounts are presented of design of potent inhibitors, largely nucleosides and acyclonucleosides, their phosphates and phosphonates, particularly of the human erythrocyte enzyme, some with Ki values in nanomolar and picomolar range, intended for induction of the immunodeficient state for clinical applications, such as prevention of host-versus-graft response in organ transplantations. Methods of assay of PNP activity are reviewed. Also described are applications of PNP from various sources as tools for the enzymatic synthesis of otherwise inaccessible therapeutic nucleoside analogues, as coupling enzymes for assays of orthophosphate in biological systems in the micromolar and submicromolar ranges, and for coupled assays of other enzyme systems.
Bzowska
A
A
Department of Biophysics, Institute of Experimental Physics, University of Warsaw, Zwirki i Wigury 93, 02-089 Warsaw, Poland. abzowska@asp.biogeo.uw.edu.pl
Kulikowska
E
E
Shugar
D
D
D000595:Amino Acid Sequence; D000818:Animals; D001681:Biological Assay / Q000379:methods; D002384:Catalysis; D015316:Genetic Therapy; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D009705:Nucleosides / Q000138:chemical synthesis / Q000627:therapeutic use; D017354:Point Mutation; D011487:Protein Conformation; D011683:Purine-Nucleoside Phosphorylase / Q000172:deficiency / Q000378:metabolism* / Q000627:therapeutic use
D009705:Nucleosides; D011683:Purine-Nucleoside Phosphorylase