Crystal structure of O-methyltransferase CalO6 from the calicheamicin biosynthetic pathway: a case of challenging structure determination at low resolution

Tsodikov, Oleg V.; Hou, Can; Walsh, Christopher T.

doi:10.1186/s12900-015-0040-6

Cited by 12 publications

(10 citation statements)

References 32 publications

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“…Methylation is an important reaction in the biosynthesis of specialized metabolites in both plants and microorganisms, where it contributes significantly to the chemical diversity of the products ( 31 , 32 ). The results presented here significantly advance our understanding of the way in which methylation shapes the branched biosynthetic pathway to the gentamicin C complex, an established drug used to combat serious infections.…”

Section: Discussionmentioning

confidence: 99%

Methyltransferases of gentamicin biosynthesis

Guo

Reva

et al. 2018

Proc. Natl. Acad. Sci. U.S.A.

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SignificanceAminoglycosides remain a vital clinical asset. Gentamicin C complex in particular is remarkably potent in treating systemic Gram-negative infections, and semisynthetic gentamicins that combat pathogen resistance or show reduced toxicity remain attractive goals. We report here the roles of clustered genes and enzymes that define a methylation network in gentamicin biosynthesis and also identify a remote gene on the chromosome encoding the essential methyltransferase GenL, which is decisive for the proportions of the five major components present in the gentamicin C complex. This is an important step toward engineered fermentation to produce single components as valuable starting materials for semisynthesis of next-generation aminoglycoside antibiotics.

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Section: Discussionmentioning

confidence: 99%

Methyltransferases of gentamicin biosynthesis

Guo

Reva

et al. 2018

Proc. Natl. Acad. Sci. U.S.A.

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“…Despite these conserved elements, EvdMO1 shows little additional sequence homology with other members of the SAM-dependent methyltransferase superfamily. [22][23][24][25][26][27][28][29][30][31][32][33] A structural homology search 34 reveals that even the most similar methyltransferases have sequence identities lower than 20% and most have an overall r.m.s. deviation greater than 2 Å when compared to EvdMΔO1 (Supplemental Table 1).…”

Section: The N-terminal Sam-dependent Methyltransferase Domain Of Evmentioning

confidence: 99%

The Structure of the Bifunctional Everninomicin Biosynthetic Enzyme EvdMO1 Suggests Independent Activity of the Fused Methyltransferase-Oxidase Domains

et al. 2018

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The orthosomycin family of natural products are decorated polysaccharides with potent antibiotic activity and complex biosynthetic pathways. The defining feature of the orthosomycins is an orthoester linkage between carbohydrate moieties that is necessary for antibiotic activity and is likely formed by a family of conserved oxygenases. Everninomicins are octasaccharide orthosomycins produced by Micromonospora carbonacea that have two orthoester linkages and a methylenedioxy bridge, three features whose formation logically require oxidative chemistry. Correspondingly, the evd gene cluster encoding Everninomicin D encodes two monofunctional nonheme iron, α-ketoglutarate dependent oxygenases and one bifunctional enzyme with an Nterminal methyltransferase domain and a C-terminal oxygenase domain. To investigate whether the activities of these domains are linked in the bifunctional enzyme EvdMO1, we determined the structure of the N-terminal methyltransferase domain to 1.1 Å and the full-length protein to 3.35 Å resolution. Both domains of EvdMO1 adopt the canonical folds of their respective superfamilies and are connected by a short linker. Each domain's active site is oriented such that it faces away from the other domain, and there is no evidence of a channel connecting the two. Our results support EvdMO1 working as a bifunctional enzyme with independent catalytic activities.

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“…Disordered regions in proteins have often been associated with function (42-45). A common characteristic of disordered regions is their ability to become ordered, typically leading to the formation of α helices, upon binding of substrates from natural targets (44).…”

Section: Discussionmentioning

confidence: 99%

New Structural Insights into the Function of the Catalytically Active Human Taspase1

Nagaratnam

Delker

Jernigan

et al. 2020

Preprint

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Proteases can play essential roles in severe human pathology, ranging from degenerative and inflammatory illnesses to infectious diseases, with some, such as Taspase1, involved in growth and progression of tumors at primary and metastatic sites. Taspase1 is a N-terminal nucleophile (Ntn)-hydrolase overexpressed in primary human cancers, coordinating cancer cell proliferation, invasion, and metastasis. Loss of Taspase1 activity disrupts proliferation of human cancer cells in vitro and in mouse xenograft models of glioblastoma, thus this protein has the potential to become a novel anticancer drug target. It belongs to the family of Ntn-hydrolases, a unique family of proteins synthesized as enzymatically inactive proenzymes that become activated upon cleavage of the peptide bond on the N-terminal side of a threonine residue, which then becomes the catalytic site nucleophile. The activation process simultaneously changes the conformation of a long domain at the C-terminus of the alpha-subunit for which no full-length structural information exists and its function is poorly understood. Here we present a novel cloning strategy to generate a fully active, circularly permuted form of Taspase1 to determine the crystallographic structure of catalytically active human Taspase1 to 3.04Å. We discovered that this region forms a long helical domain and is indispensable for the catalytic activity of Taspase1. Together, our study highlights the importance of this element for the enzymatic activity of Ntn-hydrolases and suggests that this long domain could be a novel target for the design of inhibitors with the potential to be developed into anticancer therapeutics.

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Crystal structure of O-methyltransferase CalO6 from the calicheamicin biosynthetic pathway: a case of challenging structure determination at low resolution

Cited by 12 publications

References 32 publications

Methyltransferases of gentamicin biosynthesis

Methyltransferases of gentamicin biosynthesis

The Structure of the Bifunctional Everninomicin Biosynthetic Enzyme EvdMO1 Suggests Independent Activity of the Fused Methyltransferase-Oxidase Domains

New Structural Insights into the Function of the Catalytically Active Human Taspase1

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