Novel intermolecular iterative mechanism for biosynthesis of mycoketide catalyzed by a bimodular polyketide synthase
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Title |
Novel intermolecular iterative mechanism for biosynthesis of mycoketide catalyzed by a bimodular polyketide synthase
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Creator |
Chopra, Tarun
Banerjee, Srijita Gupta, Sarika Yadav, Gitanjali Anand, Swadha Surolia, Avadhesha Roy, Rajendra P. Mohanty, Debasisa Gokhale, Rajesh S. |
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Subject |
Polyketide synthases
Mycobacterium tuberculosis Mycoketide Catalyzed Biosynthesis Intermolecular Iterative Mechanism |
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Description |
In recent years, remarkable versatility of polyketide synthases (PKSs) has been recognized; both in terms of their structural and functional organization as well as their ability to produce compounds other than typical secondary metabolites. Multifunctional Type I PKSs catalyze the biosynthesis of polyketide products by either using the same active sites repetitively (iterative) or by using these catalytic domains only once (modular) during the entire biosynthetic process. The largest open reading frame in Mycobacterium tuberculosis, pks12, was recently proposed to be involved in the biosynthesis of mannosyl-b-1-phosphomycoketide (MPM). The PKS12 protein contains two complete sets of modules and has been suggested to synthesize mycoketide by five alternating condensations of methylmalonyl and malonyl units by using an iterative mode of catalysis. The bimodular iterative catalysis would require transfer of intermediate chains from acyl carrier protein domain of module 2 to ketosynthase domain of module 1. Such bimodular iterations during PKS biosynthesis have not been characterized and appear unlikely based on recent understanding of the three-dimensional organization of these proteins. Moreover, all known examples of iterative PKSs so far characterized involve unimodular iterations. Based on cell-free reconstitution of PKS12 enzymatic machinery, in this study, we provide the first evidence for a novel ‘‘modularly iterative’’ mechanism of biosynthesis. By combination of biochemical, computational, mutagenic, analytical ultracentrifugation and atomic force microscopy studies, we propose that PKS12 protein is organized as a large supramolecular assembly mediated through specific interactions between the C- and N-terminus linkers. PKS12 protein thus forms a modular assembly to perform repetitive condensations analogous to iterative proteins. This novel intermolecular iterative biosynthetic mechanism provides new perspective to our understanding of polyketide biosynthetic machinery and also suggests new ways to engineer polyketide metabolites. The characterization of novel molecular mechanisms involved in biosynthesis of mycobacterial virulent lipids has opened new avenues for drug discovery. RSG is a HHMI International Research Scholar and is also supported by Swarnajayanti Fellowship from DST, India. This work was also supported by the Centre of Excellence grant from DBT, India to RSG and AS. |
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Date |
2013-11-07T05:15:42Z
2013-11-07T05:15:42Z 2008 27 May 2008 |
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Type |
Article
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Identifier |
PLoS Biol., 6(7): e163
http://hdl.handle.net/123456789/83 |
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Language |
en
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Publisher |
PLOS
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