Energetic mapping of transition state analogue interactions with human and Plasmodium falciparum purine nucleoside phosphorylases

Andrzej Lewandowicz, Erika A Taylor Ringia, Li Min Ting, Kami Kim, Peter C. Tyler, Gary B. Evans, Olga V. Zubkova, Simon Mee, Gavin F. Painter, Dirk H. Lenz, Richard H. Furneaux, Vern L. Schramm

Research output: Contribution to journalArticle

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Abstract

Human purine nucleoside phosphorylase (huPNP) is essential for human T-cell division by removing deoxyguanosine and preventing dGTP imbalance. Plasmodium falciparum expresses a distinct PNP (PfPNP) with a unique substrate specificity that includes 5′-methylthioinosine. The PfPNP functions both in purine salvage and in recycling purine groups from the polyamine synthetic pathway. Immucillin-H is an inhibitor of both huPNP and PfPNPs. It kills activated human T-cells and induces purine-less death in P. falciparum. Immucillin-H is a transition state analogue designed to mimic the early transition state of bovine PNP. The DADMe-Immucillins are second generation transition state analogues designed to match the fully dissociated transition states of huPNP and PfPNP. Immucillins, DADMe-Immucillins and related analogues are compared for their energetic interactions with human and P. falciparum PNPs. Immucillin-H and DADMe-Immucillin-H are 860 and 500 pM inhibitors against P. falciparum PNP but bind human PNP 15-35 times more tightly. This common pattern is a result of kcat for huPNP being 18-fold greater than kcat for PfPNP. This energetic binding difference between huPNP and PfPNP supports the k chem/kcat binding argument for transition state analogues. Preferential PfPNP inhibition is gained in the Immucillins by 5′-methylthio substitution which exploits the unique substrate specificity of PfPNP. Human PNP achieves part of its catalytic potential from 5′-OH neighboring group participation. When PfPNP acts on 5′-methylthioinosine, this interaction is not possible. Compensation for the 5′-OH effect in the P. falciparum enzyme is provided by improved leaving group interactions with Asp206 as a general acid compared with Asn at this position in huPNP. Specific atomic modifications in the transition state analogues cause disproportionate binding differences between huPNP and PfPNPs and pinpoint energetic binding differences despite similar transition states.

Original languageEnglish (US)
Pages (from-to)30320-30328
Number of pages9
JournalJournal of Biological Chemistry
Volume280
Issue number34
DOIs
StatePublished - Aug 26 2005

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Purine-Nucleoside Phosphorylase
Plasmodium falciparum
T-cells
Salvaging
Deoxyguanosine
Substrate Specificity
Polyamines
Substrates
Recycling
Substitution reactions
T-Lymphocytes
Acids
Cell Division
Enzymes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Energetic mapping of transition state analogue interactions with human and Plasmodium falciparum purine nucleoside phosphorylases. / Lewandowicz, Andrzej; Ringia, Erika A Taylor; Ting, Li Min; Kim, Kami; Tyler, Peter C.; Evans, Gary B.; Zubkova, Olga V.; Mee, Simon; Painter, Gavin F.; Lenz, Dirk H.; Furneaux, Richard H.; Schramm, Vern L.

In: Journal of Biological Chemistry, Vol. 280, No. 34, 26.08.2005, p. 30320-30328.

Research output: Contribution to journalArticle

Lewandowicz, A, Ringia, EAT, Ting, LM, Kim, K, Tyler, PC, Evans, GB, Zubkova, OV, Mee, S, Painter, GF, Lenz, DH, Furneaux, RH & Schramm, VL 2005, 'Energetic mapping of transition state analogue interactions with human and Plasmodium falciparum purine nucleoside phosphorylases', Journal of Biological Chemistry, vol. 280, no. 34, pp. 30320-30328. https://doi.org/10.1074/jbc.M505033200
Lewandowicz, Andrzej ; Ringia, Erika A Taylor ; Ting, Li Min ; Kim, Kami ; Tyler, Peter C. ; Evans, Gary B. ; Zubkova, Olga V. ; Mee, Simon ; Painter, Gavin F. ; Lenz, Dirk H. ; Furneaux, Richard H. ; Schramm, Vern L. / Energetic mapping of transition state analogue interactions with human and Plasmodium falciparum purine nucleoside phosphorylases. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 34. pp. 30320-30328.
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