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. Author manuscript; available in PMC: 2011 Sep 21.
Published in final edited form as: Biochemistry. 2010 Sep 21;49(37):7978–7987. doi: 10.1021/bi101056m

Figure 5.

Figure 5

Hydrolysis of 300 μM GD. All reactions were conducted in a volume of 200 μL and initiated by the injection of 6 μL of GD into the enzyme solution. Hydrolysis of GD (panels A, C, E, and G) was followed by monitoring the heat liberated as a function of time. Following each experiment the unreacted fraction of GD was analyzed by gas chromatography with a chiral separation column (panels B, D, F, and H). Order of elution from the gas chromatography column is SPRC (13 min), RPRC (14.1 min), SPSC (14.5 min), RPSC (15 min). (A) Hydrolysis of GD by 800 nM wild-type PTE as a function of time. Line is fit to single exponential. (B) Analysis of GD remaining following reaction with wild-type PTE. (C) Hydrolysis of GD by 200 nM G60A as a function of time. Line is a fit to the sum of 2 exponentials. (D) Analysis of unreacted GD following reaction with G60A. (E) Hydrolysis of GD by 600 nM H254G/H257W/L303T as a function of time. The line represents a fit of the data to a single exponential. (F) Analysis of uneacted GD remaining following reaction with H254G/H257W/L303T. (G) Hydrolysis of GD by100 nM H257Y/L303T as function of time. The line is a fit to the sum of two exponentials. (H) Analysis of unreacted GD remaining following reaction with H257Y/L303T.