Amino acid sequence of Escherichia coli citrate synthase

Bhayana, Vipin; Duckworth, Harry W.

doi:10.1021/bi00308a008

Cited by 37 publications

(27 citation statements)

References 28 publications

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“…22) No homologies could be found between the N-terminal amino acid sequences of citrate synthases of E. coli and pig heart and that of 469 citrate synthase purified from S. hygroscopicus in this study. This may be accounted for by the following assumptions, (1) the gene of S. hygroscopicus coding for citrate synthase may be of evolutionarily different origin from that of E. coli or pig heart, or (2) since only 20 Nterminal aminoacids of S. hygroscopicus enzyme was identified, no homology could be found among the citrate synthases of different origin.…”

Section: Aminoacid Sequence Of N-terminal Portioncontrasting

confidence: 54%

Studies on the biosynthesis of bialaphos (SF-1293). Part X. Purification and characterization of citrate synthase from Streptomyces hygroscopicus SF-1293 and comparison of its properties with those of 2-phosphinomethylmalic acid synthase.

Shimotohno

Imai

Murakami

et al. 1990

Agricultural and Biological Chemistry

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Section: Aminoacid Sequence Of N-terminal Portioncontrasting

confidence: 54%

Studies on the biosynthesis of bialaphos (SF-1293). Part X. Purification and characterization of citrate synthase from Streptomyces hygroscopicus SF-1293 and comparison of its properties with those of 2-phosphinomethylmalic acid synthase.

Shimotohno

Imai

Murakami

et al. 1990

Agricultural and Biological Chemistry

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“…The amino acid sequences of both E. coli citrate synthase and the enzyme isolated from pig heart have been determined (3,4). These enzymes share significant homology, especially in areas (identified by X-ray crystallographic studies of the pig heart enzyme) that contain residues involved in substrate binding (3). Comparison of the deduced rickettsial enzyme sequence to those of both the pig heart and E. coli enzymes (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…In addition, the two types differ in their binding affinity for the substrate acetyl coenzyme A (acetyl-CoA) (2, 7, 24). Interestingly, the mnonomers of both the large and small enzymes are comparable in size and share amino acid sequence homology (3,4,22,41). The amino acid sequences of the pig heart (4) and Escherichia coli (3) enzymes have been determined, as have the DNA sequences of the Saccharomyces cerevisiae (37) and E. coli (22) pig heart enzyme by X-ray crystallographic techniques has provided detailed molecular structure information and active site identification (27,43).…”

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confidence: 99%

Nucleotide sequence of the Rickettsia prowazekii citrate synthase gene

Wood¹,

Williamson²,

Winkler³

et al. 1987

J Bacteriol

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The Rickettsia prowazekii citrate synthase (gitA) gene, previously cloned in Escherichia coli, was localized to a 2.0-kilobase chromosomnal fragment. DNA sequence analysis of a portion of this fragment revealed an open reading frame of 1,308 base pafrs that encodes a protein of 435 amino acids with a molecular weight of 49,171. This translation product is comparable in size to both the E. coli Snd pig heart citrate synthase monomers and to the protein synthesized in E. coli minicells containing the rickettsial gene. Comparisons between the deduced amino acid sequence of R. prowazekii citrate synthase and those of the E. coli and pig heart enzymes revealed extensive homology (59%) between the two bacterial proteins. In contrast, only 20% of the rickettsial enzyme residues were shared with the functionally similar pig heart enzyine residUes. Upstream from the open reading frame and in close proximity to one anothet, sequences with homology to E. coli consensus sequences for RNA polymerase and ribosome binding were identified. S1 nuclease mapping experiments demobstrated that the start of transcription for this gene in E. coli was located in the upstream region. Codon usage in the rickettsial git4 gene was found to be very biased and differed from the pattern observed in E. coli. Adenine and uracil were used preferentially in the third base position of rickettsial codons.Rickettsia prowazekii is an obligate intracellular parasitic bacterium that multiplies within the cytoplasm of its eucaryotic host cell rather than within a phagosome or phagolysosome. Its exploitation of this environment is reflected in a variety of novel features, most notably the existence of an ADP-ATP transport system that permits the rickettsiae to accumulate ATP directly from the cytoplasm of the host cell (44). However, the rickettsiae are not strict energy parasites and can generate their own ATP via the tricarboxylic acid cycle and oxidative phosphorylation (39). This ability is probably necessary for cell viability during periods when the host cell ATP reserves are depleted or during transit of the bacterium to a new host cell. Central to the regulation of the tricarboxylic acid cycle is the enzyme citrate synthase.Citrate synthase, the first enzyme of the tricarboxylic acid cycle, has been extensively studied in a variety of procaryotic and. eucaryotic cells (30,, 41, 43). These studies have identified two main types of citrate synthase: a "small" type (molecular weight, -106,000) found in eucaryotic cells and gram-positive bacteria and a "large" type (molecular weight, -250,000) found exclusively in gram-negative bacteria. The small enzyme is a dimer composed of two identical subunits, while the large enzyme is a multimer composed of four to six identical subunits. The two types of enzyme also differ in their sensitivity to certain effectors. The small enzyme is senlsitive in vitro to inhibition by ATP but not by at-ketoglutarate or NADH, while the large enzyme is sensitive in vitro to inhibition by a-ketoglutarate and NADH but not by ...

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“…Escherichia coli and Acinetobacter anitratum citrate synthases are strongly homologous in amino acid sequence and more distantly resemble the nonallosteric citrate synthase of eucaryotes (2,8,24). Pseudomonas citrate synthases are also allosteric, and studies of partially purified preparations suggest that they have kinetic properties intermediate between those of the E. coli and A. anitratum enzymes (11,14,15, 20).…”

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confidence: 99%

Cloning, sequencing, and expression of the gene for NADH-sensitive citrate synthase of Pseudomonas aeruginosa

1989

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The structural gene for the alosteric citrate syntbase of Pseudomonas aeruginosa has been dlon fom a genomic lbrary by using the Escherichia coi citrate synthase gene as a hybridiztion probe under of reduced s ncy. Subcloing of portions of the origial 10-obe-pair (kbp) clone led to isn of the structural gene, with its promoter, witin a 2,083-bp length of DNA flanked by sites for KpnI and BanII. The nucleotide sequence of this fragment is presented; the inferred amino acid sequence was 70 and 76% id , respectively, with the itrate synthase seqces from E. co1i and Acinetobacter anitsutu, two other aeruginosa and isolates of P. putida, P. stutzeri, and P. aicalgenes. When crude extracts of eaci of these four isolates were mixed with antiserum raised against purified P. aeruginosa citrate synthase, however, only the P.akaligenes extract cross-reacted.The citrate synthases of gram-negative bacteria are allosteric enzymes, whose activity is inhibited strongly and specifically by NADH (35). Escherichia coli and Acinetobacter anitratum citrate synthases are strongly homologous in amino acid sequence and more distantly resemble the nonallosteric citrate synthase of eucaryotes (2,8,24). Pseudomonas citrate synthases are also allosteric, and studies of partially purified preparations suggest that they have kinetic properties intermediate between those of the E. coli and A. anitratum enzymes (11,14,15, 20). Until now, no one has succeeded in obtaining a pure sample of a Pseudomonas citrate synthase which is still inhibited by NADH, perhaps because the enzyme is sensitive to attack by proteases present in crude extracts of this organism. The picture has been further complicated by the demonstration that two peaks of citrate synthase activity are obtained when crude extracts of various pseudomonads are chromatographed; it has been argued that these peaks represent two distinct citrate synthases, one allosteric and the other not (22,31).In this paper we report the molecular cloning and DNA sequencing of the structural gene for the NADH-sensitive citrate synthase of an isolate ofPseudomonas aeruginosa. A plasmid subclone, harbored in an E. coli host which itself lacks citrate synthase protein, produces moderate amounts of Pseudomonas enzyme, which upon purification retains much of its sensitivity to NADH. Unexpectedly, three peaks of enzyme activity were obtained from cation-exchange chromatography, which were similar in kinetic properties and must all have arisen from the same cloned gene.MATERIALS

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Amino acid sequence of Escherichia coli citrate synthase

Cited by 37 publications

References 28 publications

Studies on the biosynthesis of bialaphos (SF-1293). Part X. Purification and characterization of citrate synthase from Streptomyces hygroscopicus SF-1293 and comparison of its properties with those of 2-phosphinomethylmalic acid synthase.

Studies on the biosynthesis of bialaphos (SF-1293). Part X. Purification and characterization of citrate synthase from Streptomyces hygroscopicus SF-1293 and comparison of its properties with those of 2-phosphinomethylmalic acid synthase.

Nucleotide sequence of the Rickettsia prowazekii citrate synthase gene

Cloning, sequencing, and expression of the gene for NADH-sensitive citrate synthase of Pseudomonas aeruginosa

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