Pre-steady-state transition-state analysis of the hydrolytic reaction catalyzed by purine nucleoside phosphorylase

Biochemistry ◽  
1995 ◽  
Vol 34 (4) ◽  
pp. 1153-1162 ◽  
Author(s):  
Paul C. Kline ◽  
Vern L. Schramm
Keyword(s):  
Steady State ◽  
Transition State ◽  
Purine Nucleoside Phosphorylase ◽  
State Transition ◽  
Purine Nucleoside ◽  
Nucleoside Phosphorylase ◽  
Hydrolytic Reaction ◽  
State Analysis

scholarly journals Catalytic-site design for inverse heavy-enzyme isotope effects in human purine nucleoside phosphorylase

2017 ◽  
Vol 114 (25) ◽  
pp. 6456-6461 ◽  
Author(s):  
Rajesh K. Harijan ◽  
Ioanna Zoi ◽  
Dimitri Antoniou ◽  
Steven D. Schwartz ◽  
Vern L. Schramm
Keyword(s):  
Steady State ◽  
Transition State ◽  
Isotope Effect ◽  
Purine Nucleoside Phosphorylase ◽  
Isotope Effects ◽  
Catalytic Site ◽  
Purine Nucleoside ◽  
Barrier Crossing ◽  
Nucleoside Phosphorylase ◽  
Inverse Effect

Heavy-enzyme isotope effects (15N-, 13C-, and 2H-labeled protein) explore mass-dependent vibrational modes linked to catalysis. Transition path-sampling (TPS) calculations have predicted femtosecond dynamic coupling at the catalytic site of human purine nucleoside phosphorylase (PNP). Coupling is observed in heavy PNPs, where slowed barrier crossing caused a normal heavy-enzyme isotope effect (kchemlight/kchemheavy > 1.0). We used TPS to design mutant F159Y PNP, predicted to improve barrier crossing for heavy F159Y PNP, an attempt to generate a rare inverse heavy-enzyme isotope effect (kchemlight/kchemheavy < 1.0). Steady-state kinetic comparison of light and heavy native PNPs to light and heavy F159Y PNPs revealed similar kinetic properties. Pre–steady-state chemistry was slowed 32-fold in F159Y PNP. Pre–steady-state chemistry compared heavy and light native and F159Y PNPs and found a normal heavy-enzyme isotope effect of 1.31 for native PNP and an inverse effect of 0.75 for F159Y PNP. Increased isotopic mass in F159Y PNP causes more efficient transition state formation. Independent validation of the inverse isotope effect for heavy F159Y PNP came from commitment to catalysis experiments. Most heavy enzymes demonstrate normal heavy-enzyme isotope effects, and F159Y PNP is a rare example of an inverse effect. Crystal structures and TPS dynamics of native and F159Y PNPs explore the catalytic-site geometry associated with these catalytic changes. Experimental validation of TPS predictions for barrier crossing establishes the connection of rapid protein dynamics and vibrational coupling to enzymatic transition state passage.

Purine Nucleoside Phosphorylase fromMycobacterium tuberculosis. Analysis of Inhibition by a Transition-State Analogue and Dissection by Parts†

Biochemistry ◽  
2001 ◽  
Vol 40 (28) ◽  
pp. 8196-8203 ◽  
Author(s):  
Luiz A. Basso ◽  
Diogenes S. Santos ◽  
Wuxian Shi ◽  
Richard H. Furneaux ◽  
Peter C. Tyler ◽  
...  
Keyword(s):  
Transition State ◽  
Purine Nucleoside Phosphorylase ◽  
Purine Nucleoside ◽  
Nucleoside Phosphorylase ◽  
Transition State Analogue

Atomic Dissection of the Hydrogen Bond Network for Transition-State Analogue Binding to Purine Nucleoside Phosphorylase†

Biochemistry ◽  
2002 ◽  
Vol 41 (49) ◽  
pp. 14489-14498 ◽  
Author(s):  
Greg A. Kicska ◽  
Peter C. Tyler ◽  
Gary B. Evans ◽  
Richard H. Furneaux ◽  
Wuxian Shi ◽  
...  
Keyword(s):  
Hydrogen Bond ◽  
Transition State ◽  
Purine Nucleoside Phosphorylase ◽  
Purine Nucleoside ◽  
Hydrogen Bond Network ◽  
Nucleoside Phosphorylase ◽  
Transition State Analogue ◽  
Bond Network

Synthesis of Transition State Analogue Inhibitors for Purine Nucleoside Phosphorylase and N-Riboside Hydrolases

Tetrahedron ◽  
2000 ◽  
Vol 56 (19) ◽  
pp. 3053-3062 ◽  
Author(s):  
Gary B. Evans ◽  
Richard H. Furneaux ◽  
Graeme J. Gainsford ◽  
Vern L. Schramm ◽  
Peter C. Tyler
Keyword(s):  
Transition State ◽  
Purine Nucleoside Phosphorylase ◽  
Purine Nucleoside ◽  
Nucleoside Phosphorylase ◽  
Transition State Analogue ◽  
Transition State Analogue Inhibitors

scholarly journals l-Enantiomers of Transition State Analogue Inhibitors Bound to Human Purine Nucleoside Phosphorylase

2008 ◽  
Vol 130 (3) ◽  
pp. 842-844 ◽  
Author(s):  
Agnes Rinaldo-Matthis ◽  
Andrew S. Murkin ◽  
Udupi A. Ramagopal ◽  
Keith Clinch ◽  
Simon P. H. Mee ◽  
...  
Keyword(s):  
Transition State ◽  
Purine Nucleoside Phosphorylase ◽  
Purine Nucleoside ◽  
Nucleoside Phosphorylase ◽  
Transition State Analogue ◽  
Transition State Analogue Inhibitors

scholarly journals Conformational States of Human Purine Nucleoside Phosphorylase at Rest, at Work, and with Transition State Analogues

Biochemistry ◽  
2010 ◽  
Vol 49 (9) ◽  
pp. 2058-2067 ◽  
Author(s):  
Achelle A. Edwards ◽  
Jeremiah D. Tipton ◽  
Michael D. Brenowitz ◽  
Mark R. Emmett ◽  
Alan G. Marshall ◽  
...  
Keyword(s):  
Transition State ◽  
Purine Nucleoside Phosphorylase ◽  
Purine Nucleoside ◽  
Conformational States ◽  
Nucleoside Phosphorylase ◽  
Transition State Analogues
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