β-ketoacyl-ACP synthase I of Escherichia coli: Nucleotide sequence of thefabB gene and identification of the cerulenin binding residue

Kauppinen, Sakari; Siggaard-Andersen, Marie-Louise; P, von Wettstein-Knowles

doi:10.1007/bf02983311

Cited by 159 publications

(127 citation statements)

References 66 publications

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“…CER Binding. The natural product CER, derived from the filamentous fungus Cephalosporium caerulens (8,27), effectively targets Cys-His-His-like KS enzymes and forms a covalent modification of the catalytic cysteine (11,12,28). CER shows a broad spectrum of antimicrobial activity (29,30), exhibits antitumor activity (31,32), and reduces food intake and body weight in mice (33).…”

Section: Fas Ks Domainmentioning

confidence: 99%

“…Several inhibitors targeting the ketoacyl synthase (KS) step of the FAS cycle have also been described, including cerulenin (CER) (8), thiolactomycin (TLM) (9), and the recently discovered platensimycin (PLM) (10). The polyketide CER inhibits both FAS type I and II KS enzymes, by covalent modification of the active site cysteine and by occupying the long acyl-binding pocket (11,12). TLM and PLM, in contrast, have been shown to be selective toward the FAS II system, blocking the malonyl and the CoA/ACP phosphopantetheinebinding sites (10,13).…”

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confidence: 99%

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Inhibition of the fungal fatty acid synthase type I multienzyme complex

Johansson

Wiltschi

Kumari

et al. 2008

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

122

134

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Fatty acids are among the major building blocks of living cells, making lipid biosynthesis a potent target for compounds with antibiotic or antineoplastic properties. We present the crystal structure of the 2.6-MDa Saccharomyces cerevisiae fatty acid synthase (FAS) multienzyme in complex with the antibiotic cerulenin, representing, to our knowledge, the first structure of an inhibited fatty acid megasynthase. Cerulenin attacks the FAS ketoacyl synthase (KS) domain, forming a covalent bond to the active site cysteine C1305. The inhibitor binding causes two significant conformational changes of the enzyme. First, phenylalanine F1646, shielding the active site, flips and allows access to the nucleophilic cysteine. Second, methionine M1251, placed in the center of the acyl-binding tunnel, rotates and unlocks the inner part of the fatty acid binding cavity. The importance of the rotational movement of the gatekeeping M1251 side chain is reflected by the cerulenin resistance and the changed product spectrum reported for S. cerevisiae strains mutated in the adjacent glycine G1250. Platensimycin and thiolactomycin are two other potent inhibitors of KSs. However, in contrast to cerulenin, they show selectivity toward the prokaryotic FAS system. Because the flipped F1646 characterizes the catalytic state accessible for platensimycin and thiolactomycin binding, we superimposed structures of inhibited bacterial enzymes onto the S. cerevisiae FAS model. Although almost all side chains involved in inhibitor binding are conserved in the FAS multienzyme, a different conformation of the loop K1413-K1423 of the KS domain might explain the observed low antifungal properties of platensimycin and thiolactomycin.cerulenin ͉ platensimycin ͉ thiolactomycin ͉ fatty acid synthesis ͉ yeast T hree different schemes for de novo synthesis of fatty acids are found in nature. Eukaryotes and advanced prokaryotes generally use the type I fatty acid synthase system (FAS I), composed of complexes of large multifunctional enzymes. Bacteria, in contrast, use the dissociated FAS II system that consists of a set of separate enzymes, each catalyzing one of the reactions of the fatty acid synthase cycle (1). A third system exists in some parasites that use membrane-bound fatty acid elongases for the synthesis of aliphatic chains (2). Despite this considerable variation, the individual reaction steps of fatty acid biosynthesis are essentially conserved in all kingdoms of life. Four basic reactions constitute a single round of elongation. In the first step, an acceptor CoA or acyl carrier protein (ACP) associated acetyl unit is condensed with malonyl-ACP to form ␤-ketobutyryl-ACP, which is subsequently reduced by an NADPH-dependent ketoacyl-ACP reductase. The resulting ␤-hydroxyacyl-ACP is dehydrated to produce enoyl-ACP and finally reduced by an enoyl reductase (ER) to form the saturated acyl-ACP, which can be further elongated in a new cycle [see supporting information (SI) Fig. S1].The importance of the fatty acid biosynthesis pathway makes the FAS sys...

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Section: Fas Ks Domainmentioning

confidence: 99%

mentioning

confidence: 99%

Inhibition of the fungal fatty acid synthase type I multienzyme complex

Johansson

Wiltschi

Kumari

et al. 2008

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

122

134

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“…Although no drugs targeting condensing enzymes are used in the clinic, two inhibitors, cerulenin (18) and thiolactomycin (19), with poor antibacterial activities were discovered more than two decades ago. Cerulenin selectively targets FabF/B forming a covalent bond with the catalytic cysteine in the active site with its tail occupying the long hydrophobic cavity that normally contains the growing acyl chain of the natural substrate (20,21). Thiolactomycin and its analogs (22,23) target the malonyl binding site of both FabH and FabF/B.…”

mentioning

confidence: 99%

Discovery of platencin, a dual FabF and FabH inhibitor with in vivo antibiotic properties

Wang

Kodali²,

Lee³

et al. 2007

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

371

366

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Emergence of bacterial resistance is a major issue for all classes of antibiotics; therefore, the identification of new classes is critically needed. Recently we reported the discovery of platensimycin by screening natural product extracts using a target-based whole-cell strategy with antisense silencing technology in concert with cell free biochemical validations. Continued screening efforts led to the discovery of platencin, a novel natural product that is chemically and biologically related but different from platensimycin. Platencin exhibits a broad-spectrum Gram-positive antibacterial activity through inhibition of fatty acid biosynthesis. It does not exhibit cross-resistance to key antibiotic resistant strains tested, including methicillin-resistant Staphylococcus aureus, vancomycin-intermediate S. aureus, and vancomycin-resistant Enterococci. Platencin shows potent in vivo efficacy without any observed toxicity. It targets two essential proteins, ␤-ketoacyl-[acyl carrier protein (ACP)] synthase II (FabF) and III (FabH) with IC 50 values of 1.95 and 3.91 g/ml, respectively, whereas platensimycin targets only FabF (IC 50 ‫؍‬ 0.13 g/ml) in S. aureus, emphasizing the fact that more antibiotics with novel structures and new modes of action can be discovered by using this antisense differential sensitivity wholecell screening paradigm.platensimycin ͉ natural product ͉ thiolactomycin ͉ antisense ͉ fatty acid synthesis

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“…This implies that either the amount or the cerulenin sensitivity of the first eluting activity must be higher than that of ~ketoacyl-ACP synthase I. Since the latter protein is abundant (8,10,12), the other cerulenin binding protein is likely to be at least as sensitive to the inhibitor. This in turn implies that neither of these two activities can be involved in the saturated fatty acid synthesis observed in the presence of 20 p.mol 9 1 t ceruleni~.…”

Section: Discussionmentioning

confidence: 99%

“…[LE392, str R, fabB '~] (17), and a derivative of DM86 termed DM86 pfabB 1 harboring a plasmid carrying the E. coli strain BfabB gene (12).…”

Section: Strains Media and Crude Protein Preparationmentioning

confidence: 99%

Role of escherichia coli β-ketoacyl-ACP synthase I in unsaturated fatty acid synthesis

Siggaard-Andersen

1988

Carlsberg Res. Commun.

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Keywords: Cerulenin, fabB, fabF, 13-ketoacyl-ACP synthase II, fatty acid synthetase l'wo activities were found in E. coli extracts which could complement unsaturated fatty acid synthesis of a cerulenin treated E. coli fatty acid synthetase. One of these is []-ketoacyl-ACP synthase I, but it is not known whether the other activity represents the previously characterized 13-ketoacyl-ACP synthase 11. A mutant strain exhibiting a temperature sensitive unsaturated fatty acid synthetic activity apparently lacked an active 13-ketoacyl-ACP synthase 1.

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β-ketoacyl-ACP synthase I of Escherichia coli: Nucleotide sequence of thefabB gene and identification of the cerulenin binding residue

Cited by 159 publications

References 66 publications

Inhibition of the fungal fatty acid synthase type I multienzyme complex

Inhibition of the fungal fatty acid synthase type I multienzyme complex

Discovery of platencin, a dual FabF and FabH inhibitor with in vivo antibiotic properties

Role of escherichia coli β-ketoacyl-ACP synthase I in unsaturated fatty acid synthesis

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