Abstract
An artificial selenoenzyme, selenolsubtilisin, was prepared by chemical conversion of the active site nucleophile (Ser 221) in the protease subtilisin into a selenocysteine. The properties of the selenol group make it a useful and general molecular probe of steric and electronic effects in catalysis involving serine (or cysteine) side chains in proteins. In the case of subtilisin, the effects of the selenium for oxygen substitution on the acylation and deacylation steps that occur during substrate hydrolysis and on partitioning of the acyl enzyme intermediate between amines and water were studied. Increases in aminolysis selectivity of up to 14,000 fold were achieved with the selenoenzyme compared to native subtilisin, suggesting that selenolsubtilisin could be useful as a peptide ligase. Like the naturally occurring selenoenzyme glutathione peroxidase, selenolsubtilisin also has interesting redox properties. It catalyzes the reduction of hydroperoxides by thiols. Comparison of the initial rates for the oxidation of 3-carboxy-4-nitro-benzenethiol by t-butyl hydroperoxide in the presence of the enzyme and a model selenocompound indicates a 17,000 fold rate enhancement due to protein binding.
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Wu, ZP., Hilvert, D. (1991). Redesign of Protein Function: A Semisynthetic Selenoenzyme. In: Gebelein, C.G. (eds) Biotechnology and Polymers. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-3844-8_26
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DOI: https://doi.org/10.1007/978-1-4615-3844-8_26
Publisher Name: Springer, Boston, MA
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