Structure of the Complex between the Antibiotic Cerulenin and Its Target, β-Ketoacyl-Acyl Carrier Protein Synthase

Moche, M.; Schneider, Günter; Edwards, Patricia C.; Dehesh, Katayoon; Lindqvist, Ylva

doi:10.1074/jbc.274.10.6031

Cited by 190 publications

(187 citation statements)

References 30 publications

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“…3 A and B and S4). A similar change of rotamer was observed in the M1251 counterpart I108 of the FabF-CER complex (34). This suggests that the acyl chain conformation of CER bound to FAS and to FabF might be related, directed away from the KS dimer interface instead of toward it, as seen in the FabB-CER complex.…”

Section: Fas Ks Domainsupporting

confidence: 54%

“…A similar main chain interaction has been suggested to stabilize the oxyanion formed during the acetyl transfer step of the condensation reaction (28). As compared with the native FAS structure (19), the active site of the KS-CER complex is in an open conformation (10,34), with the phenyl group of F1646 flipped ( Fig. 3 A and B).…”

Section: Fas Ks Domainmentioning

confidence: 89%

“…3 A and B). This conformation provides free access to the nucleophilic cysteine and enables CER to mimic the acyl-enzyme intermediate of the condensation reaction (34).…”

Section: Fas Ks Domainmentioning

confidence: 99%

“…A superposition of two bacterial type II KS-CER complex structures (28,34) onto the FAS condensing enzyme showed that the amide half of the inhibitor colocalizes with the 4 to 5 F o Ϫ F c density peaks in the different KS active sites of the FAS particle. As was shown for the CER-inhibited FabF/FabB enzymes, the C2 atom of CER formed a covalent bond with the SH group of the catalytic cysteine, whereas the CER amide was located close to the two active site histidine residues, H1542 and H1583.…”

Section: Fas Ks Domainmentioning

confidence: 99%

“…This suggests that the acyl chain conformation of CER bound to FAS and to FabF might be related, directed away from the KS dimer interface instead of toward it, as seen in the FabB-CER complex. The FabF I108 has been proposed to be responsible for the low CER sensitivity compared to FabB, which has a glycine residue at this position (28,34). Because the IC 50 value of FAS CER inhibition (35) is in the range of FabB, the FAS KS has to be able to compensate for the rearrangement of the methionine, for example, by stacking interactions to the CER C7-C8 double bond and a better fit of the rigid CER tail to the end of the acyl-binding pocket.…”

Section: Fas Ks Domainmentioning

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.

118

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 Domainsupporting

confidence: 54%

Section: Fas Ks Domainmentioning

confidence: 89%

“…3 A and B). This conformation provides free access to the nucleophilic cysteine and enables CER to mimic the acyl-enzyme intermediate of the condensation reaction (34).…”

Section: Fas Ks Domainmentioning

confidence: 99%

Section: Fas Ks Domainmentioning

confidence: 99%

Section: Fas Ks Domainmentioning

confidence: 99%

See 3 more Smart Citations

Inhibition of the fungal fatty acid synthase type I multienzyme complex

Johansson

Wiltschi

Kumari

et al. 2008

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

118

134

View full text Add to dashboard Cite

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The Molecular Basis of Catalysis by SDR Family Members Ketoacyl‐ACP Reductase FabG and Enoyl‐ACP Reductase FabI in Type‐II Fatty Acid Biosynthesis

Zhou,

Zhang,

Wang

et al. 2023

Angewandte Chemie

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The short‐chain dehydrogenase/reductase (SDR) superfamily members acyl‐ACP reductases FabG and FabI are indispensable core enzymatic modules and catalytic orientation controllers in type‐II fatty acid biosynthesis. Herein, we report their distinct substrate allosteric recognition and enantioselective reduction mechanisms. FabG achieves allosteric regulation of ACP and NADPH through ACP binding across two adjacent FabG monomers, while FabI follows an irreversible compulsory order of substrate binding in that NADH binding must precede that of ACP on a discrete FabI monomer. Moreover, FabG and FabI utilize a backdoor residue Phe187 or a “rheostat” α8 helix for acyl chain length selection, and their corresponding triad residues Ser142 or Tyr145 recognize the keto‐ or enoyl‐acyl substrates, respectively, facilitating initiation of nucleophilic attack by NAD(P)H. The other two triad residues (Tyr and Lys) mediate subsequent proton transfer and (R)‐3‐hydroxyacyl‐ or saturated acyl‐ACP production.

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