Inhibition of glyceraldehyde-3-phosphate dehydrogenase by pentalenolactone: Kinetic and mechanistic studies

Cane, David E.; Sohng, Jae Kyung

doi:10.1016/0003-9861(89)90006-4

Cited by 41 publications

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“…In addition, it has been shown to inhibit viral replication of herpes simplex virus 1 and herpes simplex virus 2 (7) and cell proliferation of rat vascular smooth muscle cells (8). Pentalenolactone inhibits glycolysis (9) by selectively inhibiting glyceraldehyde-3-phosphate dehydrogenase (10). The antibiotic was shown to irreversibly inactivate glyceraldehyde-3-phosphate dehydrogenase by alkylation of the active site cysteine residue (11).…”

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Crystal Structure of the Non-heme Iron Dioxygenase PtlH in Pentalenolactone Biosynthesis

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Ōmura

Ikeda

et al. 2007

Journal of Biological Chemistry

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The non-heme iron dioxygenase PtlH from the soil organism Streptomyces avermitilis is a member of the iron(II)/␣-ketoglutarate-dependent dioxygenase superfamily and catalyzes an essential reaction in the biosynthesis of the sesquiterpenoid antibiotic pentalenolactone. To investigate the structural basis for substrate recognition and catalysis, we have determined the x-ray crystal structure of PtlH in several complexes with the cofactors iron, ␣-ketoglutarate, and the non-reactive enantiomer of the substrate, ent-1-deoxypentalenic acid, in four different crystal forms to up to 1.31 Å resolution. The overall structure of PtlH forms a double-stranded barrel helix fold, and the cofactor-binding site for iron and ␣-ketoglutarate is similar to other double-stranded barrel helix fold enzymes. Additional secondary structure elements that contribute to the substrate-binding site in PtlH are not conserved in other double-stranded barrel helix fold enzymes. Binding of the substrate enantiomer induces a reorganization of the monoclinic crystal lattice leading to a disorder-order transition of a C-terminal ␣-helix. The newly formed helix blocks the major access to the active site and effectively traps the bound substrate. Kinetic analysis of wild type and site-directed mutant proteins confirms a critical function of two arginine residues in substrate binding, while simulated docking of the enzymatic reaction product reveals the likely orientation of bound substrate.Streptomyces avermitilis is a Gram-positive soil-dwelling organism that is used industrially for the production of the widely used anthelminthic avermectins. Recently, an S. avermitilis gene cluster for the biosynthesis of the sesquiterpenoid antibiotic pentalenolactone ( Fig. 1A) has been characterized (1). Pentalenolactone was first identified from Streptomyces roseogriseus (2) and is also produced by several Streptomyces strains (3-5). The antibiotic is active against Gram-positive and Gram-negative bacteria as well as fungi and protozoa (6). In addition, it has been shown to inhibit viral replication of herpes simplex virus 1 and herpes simplex virus 2 (7) and cell proliferation of rat vascular smooth muscle cells (8). Pentalenolactone inhibits glycolysis (9) by selectively inhibiting glyceraldehyde-3-phosphate dehydrogenase (10). The antibiotic was shown to irreversibly inactivate glyceraldehyde-3-phosphate dehydrogenase by alkylation of the active site cysteine residue (11). An inducible, pentalenolactone-insensitive glyceraldehyde-3-phosphate dehydrogenase isozyme that is part of the biosynthetic gene cluster confers self-resistance against the antibiotic in S. avermitilis and other Streptomyces strains (1,(12)(13)(14).We recently reported the molecular cloning and biochemical characterization of PtlH, an essential non-heme iron-dependent dioxygenase in the pentalenolactone biosynthesis pathway (15). The enzyme catalyzes the Fe(II)-and ␣-ketoglutarate-dependent hydroxylation of 1-deoxypentalenic acid to 11␤-hydroxy-1-deoxypentalenic acid (Fig. 1B). Non-heme...

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Crystal Structure of the Non-heme Iron Dioxygenase PtlH in Pentalenolactone Biosynthesis

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Ōmura

Ikeda

et al. 2007

Journal of Biological Chemistry

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“…The compound was first isolated from Streptomyces roseogriseus 1957 and later from over 30 Streptomyces species. This isoprene-derived natural product exhibits various biological activities which include diverse antibacterial and antifungal activity, as well as potent inhibitory activity toward the glycolytic enzyme glyceraldehyde 3-phosphate dehydrogenase [8]. It was also reported to inhibit the replication of DNA viruses such as HSV-1 and HSV-2 [37].…”

Section: Pentalenolactonementioning

confidence: 99%

Identification and activation of novel biosynthetic gene clusters by genome mining in the kirromycin producer Streptomyces collinus Tü 365

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Kulik

Härtner

et al. 2016

Journal of Industrial Microbiology and Biotechnology

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Streptomycetes are prolific sources of novel biologically active secondary metabolites with pharmaceutical potential. S. collinus Tü 365 is a Streptomyces strain, isolated 1972 from Kouroussa (Guinea). It is best known as producer of the antibiotic kirromycin, an inhibitor of the protein biosynthesis interacting with elongation factor EF-Tu. Genome Mining revealed 32 gene clusters encoding the biosynthesis of diverse secondary metabolites in the genome of Streptomyces collinus Tü 365, indicating an enormous biosynthetic potential of this strain. The structural diversity of secondary metabolisms predicted for S. collinus Tü 365 includes PKS, NRPS, PKS-NRPS hybrids, a lanthipeptide, terpenes and siderophores. While some of these gene clusters were found to contain genes related to known secondary metabolites, which also could be detected in HPLC-MS analyses, most of the uncharacterized gene clusters are not expressed under standard laboratory conditions. With this study we aimed to characterize the genome information of S. collinus Tü 365 to make use of gene clusters, which previously have not been described for this strain. We were able to connect the gene clusters of a lanthipeptide, a carotenoid, five terpenoid compounds, an ectoine, a siderophore and a spore pigment-associated gene cluster to their respective biosynthesis products.

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“…Keywords farnesyl diphosphate; pentalenolactone; Streptomyces; biosynthesis; GC-MS; proton NMR; COSY; dehydrogenase Pentalenolactone (1) is a sesquiterpenoid antibiotic that has been isolated from over 30 Streptomyces species. The antibiotic, which is active against a variety of microorganisms including Gram-positive and Gram-negative bacteria, fungi and protozoa, irreversibly inactivates the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase [1,2]. Pentalenolactone also inhibits the replication of DNA viruses such as HSV-1 and HSV-2 [3] as well as smooth muscle cell proliferation [4].…”

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“…Deletion of this entire operon from S. avermitilis abolished pentalenolactone formation, while transfer of the intact 13.4 kb cluster to S. lividans 1326 resulted in formation of pentalenic acid, a known pentalenolactone shunt product [9]. Heterologous expression of the ptlA gene (SAV2998) in Escherichia coli and analysis of the resultant protein established that PtlA is a pentalenene synthase that catalyzes the cyclization of farnesyl diphosphate (2) to the sesquiterpene hydrocarbon pentalenene (3) [9], as predicted by the 76% identity of the PtlA gene product to the well characterized pentalenene synthase of Streptomyces UC5319 [10]. The gap1 gene (SAV2990) at the 5′-end of the cluster was shown to encode a pentalenolactone-insensitive glyceraldehyde-3-phosphate dehydrogenase corresponding to the pentalenolactone resistance gene [9].…”

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Pentalenolactone biosynthesis: Molecular cloning and assignment of biochemical function to PtlF, a short-chain dehydrogenase from Streptomyces avermitilis, and identification of a new biosynthetic intermediate

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Ōmura

Ikeda

et al. 2007

Archives of Biochemistry and Biophysics

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Pentalenolactone (1) is an antibiotic that has been isolated from many species of Streptomyces. The putative dehydrogenase encoded by the ptlF gene (SAV2993) found within the Streptomyces avermitilis pentalenolactone gene cluster was cloned and overexpressed in Escherichia coli. PtlF, which belongs to the short-chain dehydrogenase/oxidoreductase superfamily, was shown to catalyze the oxidation of 1-deoxy-11β-hydroxypentalenic acid (9) to 1-deoxy-11-oxopentalenic acid (10), a new intermediate of the pentalenolactone biosynthetic pathway. The methyl ester of 10 was characterized by NMR, GC-MS and high resolution mass spectrometry. PtlF exhibited a 150-fold preference for β-NAD + over β-NADP + . PtlF had a pH optimum of 8.0 in the physiological pH range, while a significant activity enhancement was observed from pH 9.0 to 11.3. At pH 8.0, PtlF had a k cat of 0.65±0.03 s −1 , with a K m for 9 of 6.5±1.5 μM and K m for NAD + of 25 ± 3 μM. Keywords farnesyl diphosphate; pentalenolactone; Streptomyces; biosynthesis; GC-MS; proton NMR; COSY; dehydrogenase Pentalenolactone (1) is a sesquiterpenoid antibiotic that has been isolated from over 30 Streptomyces species. The antibiotic, which is active against a variety of microorganisms including Gram-positive and Gram-negative bacteria, fungi and protozoa, irreversibly inactivates the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase [1,2]. Pentalenolactone also inhibits the replication of DNA viruses such as HSV-1 and HSV-2 [3] as well as smooth muscle cell proliferation [4]. Isotopic incorporation experiments using intact cultures of Streptomyces UC5319 have firmly established that pentalenene (3), which is formed by the cyclization of farnesyl diphosphate (2), is the parent hydrocarbon of the pentalenolactone family of metabolites [5]. A variety of oxidized metabolites that are plausible intermediates in the conversion of pentalenene to pentalenolactone have been isolated, including 1-1 Corresponding author: Fax: +1-401-863-9368; E-mail: David_Cane@brown.edu.Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. (4)) and ptlH (SAV2991), a non-heme iron/α-ketoglutarate-dependent hydroxylase, that we have shown catalyzes the oxidative conversion of 1-deoxypentalenic acid (4) to 1-deoxy-11β-hydroxypentalenic acid (9) [12]. Of the remaining ORFs, ptlB corresponds to a typical farnesyl diphosphate synthase, while ptlR is a putative transcriptional regulator and PtlG represents a likely transmembrane efflux protein [10]. Finally, ptlF (SAV2993) appears to encode a typical short-chain dehydrogenase/reductase [10]. As illustrate...

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Inhibition of glyceraldehyde-3-phosphate dehydrogenase by pentalenolactone: Kinetic and mechanistic studies

Cited by 41 publications

References 35 publications

Crystal Structure of the Non-heme Iron Dioxygenase PtlH in Pentalenolactone Biosynthesis

Crystal Structure of the Non-heme Iron Dioxygenase PtlH in Pentalenolactone Biosynthesis

Identification and activation of novel biosynthetic gene clusters by genome mining in the kirromycin producer Streptomyces collinus Tü 365

Pentalenolactone biosynthesis: Molecular cloning and assignment of biochemical function to PtlF, a short-chain dehydrogenase from Streptomyces avermitilis, and identification of a new biosynthetic intermediate

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