The 1.8 Å crystal structure of the dimeric peroxisomal 3-ketoacyl-CoA thiolase of Saccharomyces cerevisiae: implications for substrate binding and reaction mechanism

Mathieu, Magali; Modis, Yorgo; Zeelen, J.P.; Engel, C.K.; Abagyan, Ruben; Ahlberg, Anders; Rasmussen, Bjarne; Lamzin, Victor S.; Kunau, Wolf‐H.; Wierenga, Rik K.

doi:10.1006/jmbi.1997.1331

Cited by 117 publications

(122 citation statements)

References 42 publications

(39 reference statements)

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“…The spFabF protein forms an ␣/␤ structure that can be divided into two halves, an N-terminal region comprising residues 1 to 250 and a C-terminal region comprising residues 251 to 409. The basic fold of the superfamily was first described for a yeast thiolase (26), and the two halves have identical core structures comprising a five-stranded mixed ␤-sheet that is flanked on one side by two ␣-helices and on the other by a single ␣-helix. A single copy of this duplicated topology was identified in the first domain of phosphoglucomutase (26).…”

Section: Resultsmentioning

confidence: 99%

“…Included in the superfamily are the thiolases involved in fatty acid degradation (␤-oxidation) (46), or sterol biosynthesis (25), and the related enzymes that synthesize polyketides in plants and bacteria (18). Structural studies on this superfamily of enzymes have revealed that they share a common ␤␣␤␣␤␣␤␤ topology which is duplicated by an internal pseudodyad that may have arisen by gene duplication (26). Members of the superfamily that have been structurally characterized include the peroxisomal 3-ketoacyl-coenzyme A (3-ketoacyl-CoA) thiolases from a variety of organisms (26), an array of bacterial fatty acidcondensing enzymes (8,19,31,36,38,40,41,45), and the polyketide chalcone synthase (13).…”

mentioning

confidence: 99%

“…Structural studies on this superfamily of enzymes have revealed that they share a common ␤␣␤␣␤␣␤␤ topology which is duplicated by an internal pseudodyad that may have arisen by gene duplication (26). Members of the superfamily that have been structurally characterized include the peroxisomal 3-ketoacyl-coenzyme A (3-ketoacyl-CoA) thiolases from a variety of organisms (26), an array of bacterial fatty acidcondensing enzymes (8,19,31,36,38,40,41,45), and the polyketide chalcone synthase (13). Based on the structural and functional differences between the enzymes, the superfamily is subdivided into three classes (31).…”

mentioning

confidence: 99%

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The 1.3-Angstrom-Resolution Crystal Structure of β-Ketoacyl-Acyl Carrier Protein Synthase II fromStreptococcus pneumoniae

Price¹,

Rock

White

2003

J Bacteriol

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The ␤-ketoacyl-acyl carrier protein synthases are members of the thiolase superfamily and are key regulators of bacterial fatty acid synthesis. As essential components of the bacterial lipid metabolic pathway, they are an attractive target for antibacterial drug discovery. We have determined the 1.3 Å resolution crystal structure of the ␤-ketoacyl-acyl carrier protein synthase II (FabF) from the pathogenic organism Streptococcus pneumoniae. The protein adopts a duplicated ␤␣␤␣␤␣␤␤ fold, which is characteristic of the thiolase superfamily. The two-fold pseudosymmetry is broken by the presence of distinct insertions in the two halves of the protein. These insertions have evolved to bind the specific substrates of this particular member of the thiolase superfamily. Docking of the pantetheine moiety of the substrate identifies the loop regions involved in substrate binding and indicates roles for specific, conserved residues in the substrate binding tunnel. The active site triad of this superfamily is present in spFabF as His 303, His 337, and Cys 164. Near the active site is an ion pair, Glu 346 and Lys 332, that is conserved in the condensing enzymes but is unusual in our structure in being stabilized by an Mg 2؉ ion which interacts with Glu 346. The active site histidines interact asymmetrically with Lys 332, whose positive charge is closer to His 303, and we propose a specific role for the lysine in polarizing the imidazole ring of this histidine. This asymmetry suggests that the two histidines have unequal roles in catalysis and provides new insights into the catalytic mechanisms of these enzymes.The ␤-ketoacyl-acyl carrier protein synthases catalyze the addition of two-carbon units onto the growing acyl chain during the elongation phase of fatty acid synthesis. This enzymatic activity involves what is an irreversible chemical event in biosynthetic pathways, the formation of a carbon-carbon bond via a Claisen-like condensation reaction (15). These condensing enzymes are members of a superfamily of enzymes whose members all catalyze essentially the same reaction. Included in the superfamily are the thiolases involved in fatty acid degradation (␤-oxidation) (46), or sterol biosynthesis (25), and the related enzymes that synthesize polyketides in plants and bacteria (18). Structural studies on this superfamily of enzymes have revealed that they share a common ␤␣␤␣␤␣␤␤ topology which is duplicated by an internal pseudodyad that may have arisen by gene duplication (26). Members of the superfamily that have been structurally characterized include the peroxisomal 3-ketoacyl-coenzyme A (3-ketoacyl-CoA) thiolases from a variety of organisms (26), an array of bacterial fatty acidcondensing enzymes (8,19,31,36,38,40,41,45), and the polyketide chalcone synthase (13). Based on the structural and functional differences between the enzymes, the superfamily is subdivided into three classes (31). The thiolases form one group, the fatty acid elongation-condensing enzymes form a second group, and those involved in polyketide synt...

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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The 1.3-Angstrom-Resolution Crystal Structure of β-Ketoacyl-Acyl Carrier Protein Synthase II fromStreptococcus pneumoniae

Price¹,

Rock

White

2003

J Bacteriol

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“…2) may be important for the interaction of the CoA moiety with DGAT, as suggested for other CoA-dependent enzymes (26,27). Hence, the alanine residue at position 232 could have a negative effect on the acyl-CoA-binding capacity of DGAT, accounting for its associated lower milk fat content.…”

Section: Discussionmentioning

confidence: 99%

Association of a lysine-232/alanine polymorphism in a bovine gene encoding acyl-CoA:diacylglycerol acyltransferase ( DGAT1 ) with variation at a quantitative trait locus for milk fat content

Winter

Krämer

Werner

et al. 2002

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

331

317

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Among the variants was a nonconservative substitution of lysine by alanine (K232A), with the lysine-encoding allele being associated with higher milk fat content. Haplotype analysis indicated the lysine variant to be ancestral. Two animals that were typed heterozygous (Qq) at the QTL based on marker-assisted QTLgenotyping were heterozygous for the K232A substitution, whereas 14 animals that are most likely qq at the QTL were homozygous for the alanine-encoding allele. An independent association study in Fleckvieh animals confirmed the positive effect of the lysine variant on milk fat content. We consider the nonconservative K232A substitution to be directly responsible for the QTL variation, although our genetic studies cannot provide formal proof.

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“…2, B and C). The reason for this is currently unknown; however, it was previously shown that two free cysteine residues that exist near the active center of thiolase are necessary for its enzymatic activity (32). The accumulated homocysteine may covalently cross-link with these cysteine residues of thiolase via disulfide bonds and inhibit its enzymatic activity.…”

Section: Fig 3 Apob100 Contents/distribution In Serum (Lipoproteinsmentioning

confidence: 99%

Abnormal Lipid Metabolism in Cystathionine β-Synthase-deficient Mice, an Animal Model for Hyperhomocysteinemia

Namekata

Enokido

Ishii

et al. 2004

Journal of Biological Chemistry

136

135

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Hyperhomocysteinemia (HHCY) is a consequence of impaired methionine/cysteine metabolism and is caused by deficiency of vitamins and/or enzymes such as cystathionine ␤-synthase (CBS). Although HHCY is an important and independent risk factor for cardiovascular diseases that are commonly associated with hepatic steatosis, the mechanism by which homocysteine promotes the development of fatty liver is poorly understood. CBS-deficient (CBS ؊/؊ ) mice were previously generated by targeted deletion of the Cbs gene and exhibit pathological features similar to HHCY patients, including endothelial dysfunction and hepatic steatosis. Here we show abnormal lipid metabolism in CBS ؊/؊ mice. Triglyceride and nonesterified fatty acid levels were markedly elevated in CBS ؊/؊ mouse liver and serum. The activity of thiolase, a key enzyme in ␤-oxidation of fatty acids, was significantly impaired in CBS ؊/؊ mouse liver. Hepatic apolipoprotein B100 levels were decreased, whereas serum apolipoprotein B100 and very low density lipoprotein levels were elevated in CBS ؊/؊ mice. Serum levels of cholesterol/ phospholipid in high density lipoprotein fractions but not of total cholesterol/phospholipid were decreased, and the activity of lecithin-cholesterol acyltransferase was severely impaired in CBS ؊/؊ mice. Abnormal high density lipoprotein particles with higher mobility in polyacrylamide gel electrophoresis were observed in serum obtained from CBS ؊/؊ mice. Moreover, serum cholesterol/ triglyceride distribution in lipoprotein fractions was altered in CBS ؊/؊ mice. These results suggest that hepatic steatosis in CBS ؊/؊ mice is caused by or associated with abnormal lipid metabolism.

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The 1.8 Å crystal structure of the dimeric peroxisomal 3-ketoacyl-CoA thiolase of Saccharomyces cerevisiae: implications for substrate binding and reaction mechanism

Cited by 117 publications

References 42 publications

The 1.3-Angstrom-Resolution Crystal Structure of β-Ketoacyl-Acyl Carrier Protein Synthase II fromStreptococcus pneumoniae

The 1.3-Angstrom-Resolution Crystal Structure of β-Ketoacyl-Acyl Carrier Protein Synthase II fromStreptococcus pneumoniae

Association of a lysine-232/alanine polymorphism in a bovine gene encoding acyl-CoA:diacylglycerol acyltransferase ( DGAT1 ) with variation at a quantitative trait locus for milk fat content

Abnormal Lipid Metabolism in Cystathionine β-Synthase-deficient Mice, an Animal Model for Hyperhomocysteinemia

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