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. Author manuscript; available in PMC: 2010 Sep 25.

Published in final edited form as: Chem Biol. 2009 Sep 25;16(9):971–979. doi: 10.1016/j.chembiol.2009.07.012

(A) Single-turnover hydrolysis reaction kinetics, plotting the concentration of hypoxanthine (isolated on a C18 analytical column by reversed-phase HPLC) formed during the hydrolysis reaction of native (), Y88G (), F159G (), H257G (), and F200G human PNP () as a function of mixing time prior to acid quenching. Data were fit to equation (1).

(B) ¹³C NMR spectra of the PNP hydrolysis reactions. [1′-¹³C]inosine (900 μM) with (A) native bovine PNP (BtPNP), (B) native human PNP (HsPNP), (C) Y88G, (D) F159G, (E) H257G, and (F) F200G. All spectra were acquired after 20 h at 25 °C with 200 μM PNP in 10% D₂O. [1] Ino, inosine; [3], d-ribose; [5] N3-Ino, N3-isoinosine.

Inline graphic — (A) Single-turnover hydrolysis reaction kinetics, plotting the concentration of hypoxanthine (isolated on a C18 analytical column by reversed-phase HPLC) formed during the hydrolysis reaction of native (), Y88G (), F159G (), H257G (), and F200G human PNP () as a function of mixing time prior to acid quenching. Data were fit to equation (1).

(B) ¹³C NMR spectra of the PNP hydrolysis reactions. [1′-¹³C]inosine (900 μM) with (A) native bovine PNP (BtPNP), (B) native human PNP (HsPNP), (C) Y88G, (D) F159G, (E) H257G, and (F) F200G. All spectra were acquired after 20 h at 25 °C with 200 μM PNP in 10% D₂O. [1] Ino, inosine; [3], d-ribose; [5] N3-Ino, N3-isoinosine.