AI Article Synopsis
- The study investigates the unique substrate binding mechanism of the taurocyamine kinase (TK) from Paragonimus westermani through site-directed mutagenesis.
- Mutations at specific amino acid positions (Y84, Y87, and A59) in both domains of TK severely affect its ability to bind and catalyze the substrate taurocyamine.
- Substituting these residues with others (like R, H, and I) leads to significant decreases in enzyme activity, highlighting their importance in substrate recognition and binding.
Article Abstract
The two-domain taurocyamine kinase (TK) from Paragonimus westermani was suggested to have a unique substrate binding mechanism. We performed site-directed mutagenesis on each domain of this TK and compared the kinetic parameters Km(Tc) and Vmax with that of the wild-type to determine putative amino acids involved in substrate recognition and binding. Replacement of Y84 on domain 1 and Y87 on domain 2 with R resulted in the loss of activity for the substrate taurocyamine. Y84E mutant has a dramatic decrease in affinity and activity for taurocyamine while Y87E has completely lost catalytic activity. Substituting H and I on the said positions also resulted in significant changes in activity. Mutation of the residues A59 on the GS region of domain 1 also caused significant decrease in affinity and activity while mutation on the equivalent position on domain 2 resulted in complete loss of activity.
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| http://dx.doi.org/10.1016/j.exppara.2013.10.008 | DOI Listing |
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