In nature, enzymes often form multienzyme complexes to enhance their catalytic efficiencies and, moreover, evolve into genetically fused multidomain enzymes. Inspired by a natural fusion cytochrome P450 (P450) containing a monooxygenase domain and a reductase domain, we have developed a heterotrimeric protein- utilized method to form a multienzyme complex composed of a bacterial P450 and its catalytically essential two redox proteins. Three distinct proliferating cell nuclear antigens (PCNAs) from Sulfolobus solfataricu s, each of which can be separately expressed, spontaneously form a heterotrimer. Fusion to the PCNAs enables complex formation of a bacterial P450 and two redox proteins through the self-assembling of the PCNAs and enhances the activity due to efficient electron transfer in the complex. This PCNAmediated multienzyme complex formation will be available for other multienzyme reactions. © Springer Science+Business Media New York 2013.
CITATION STYLE
Hirakawa, H., & Nagamune, T. (2013). Use of sulfolobus solfataricus PCNA subunit proteins to direct the assembly of multimeric enzyme complexes. Methods in Molecular Biology, 978, 149–163. https://doi.org/10.1007/978-1-62703-293-3_11
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