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. 2011 Nov 8;40(5):2065–2075. doi: 10.1093/nar/gkr905

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© The Author(s) 2011. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 8. — His¹⁶⁷ is a key amino acid determining NExo activity. (A–C) Comparison of the phosphatase activity of NExo, NExo His¹⁶⁷Gly and NExo His¹⁶⁷Ser were performed by incubating 3 nM enzyme with 150 nM substrate 20-PO₄ under phosphatase assay conditions. Aliquots were removed at specified times, quenched and separated by 20% denaturing PAGE; representative lanes are shown for clarity. (D) Analysis of the full reaction profile is shown with fits to straight line plots to determine the initial velocity for NExo (solid circles, solid line; k = 0.15/s); NExo His¹⁶⁷Gly (open triangles, dashed line; k = 0.022/s) and NExo His¹⁶⁷Ser (closed triangles, dotted line; k = 0.023/s).