Ronald G. Duggleby

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Sulfonation is an important reaction in the metabolism of numerous xenobiotics, drugs, and endogenous compounds. A supergene family of enzymes called sulfotransferases (SULTs) catalyze this reaction. In most cases, the addition of a sulfonate moiety to a compound increases its water solubility and decreases its biological activity. However, many of these(More)
MOTIVATION The derivation of steady-state equations is frequently carried out in enzyme kinetic studies. Done manually, this becomes tedious and prone to human error. The computer programs now available which are able to accept reaction mechanisms of some complexity are focused only on the strict steady-state approach. RESULTS Here we present a computer(More)
BACKGROUND Site-directed mutagenesis is an efficient method to alter the structure and function of genes. Here we report a rapid and efficient megaprimer-based polymerase chain reaction (PCR) mutagenesis strategy that by-passes any intermediate purification of DNA between two rounds of PCR. RESULTS The strategy relies on the use of a limiting(More)
Despite the number of computer-assisted methods described for the derivation of steady-state equations of enzyme systems , most of them are focused on strict steady-state conditions or are not able to solve complex reaction mechanisms. Moreover, many of them are based on computer programs that are either not readily available or have limitations. We present(More)
Acetohydroxyacid synthase (AHAS; EC catalyzes the first common step in branched-chain amino acid biosynthesis. The enzyme is inhibited by several chemical classes of compounds and this inhibition is the basis of action of the sulfonylurea and imidazolinone herbicides. The commercial sulfonylureas contain a pyrimidine or a triazine ring that is(More)
Ketol-acid reductoisomerase (EC catalyses the second reaction in the biosynthesis of branched-chain amino acids. The reaction involves an Mg2+ -dependent alkyl migration followed by an NADPH-dependent reduction of the 2-keto group. Here, the crystallization of the Escherichia coli enzyme is reported. A form with a C-terminal hexahistidine tag(More)
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