Coenzyme-A-binding site of chloramphenicol acetyltransferase

1988 ◽  
Vol 16 (5) ◽  
pp. 715-716 ◽  
Author(s):  
PHILIP J. DAY ◽  
ANN LEWENDON ◽  
WILLIAM V. SHAW
Keyword(s):  
Binding Site ◽  
Coenzyme A ◽  
Chloramphenicol Acetyltransferase

pGR71 plasmid promotor sequence temporally regulated in Bacillus subtilis

1998 ◽  
Vol 44 (12) ◽  
pp. 1186-1192
Author(s):  
Guy Daxhelet ◽  
Philippe Gilot ◽  
Etienne Nyssen ◽  
Philippe Hoet
Keyword(s):  
Bacillus Subtilis ◽  
Binding Site ◽  
Transcription Initiation ◽  
Promoter Sequence ◽  
Initiation Site ◽  
Chloramphenicol Acetyltransferase ◽  
Ribosome Binding Site ◽  
Chloramphenicol Resistance ◽  
E Coli ◽  
Ribosome Binding

pGR71, a composite of plasmids pUB110 and pBR322, replicates in Escherichia coli and in Bacillus subtilis. It carries the chloramphenicol resistance gene (cat) from Tn9, which is not transcribed in either host by lack of a promoter. The cat gene is preceded by a Shine-Dalgarno sequence functional in E. coli but not in B. subtilis. Deleted pGR71 plasmids were obtained in B. subtilis when cloning foreign viral DNA upstream of this cat sequence, as well as by BAL31 exonuclease deletions extending upstream from the cat into the pUB110 moiety. These mutant plasmids expressed chloramphenicol acetyltransferase (CAT), conferring on B. subtilis resistance to high chloramphenicol concentrations. CAT expression peaked at the early postexponential phase of B. subtilis growth. The transcription initiation site of cat, determined by primer extension, was located downstream of a putative promoter sequence within the pUB110 moiety. N-terminal amino acid sequencing showed that native CAT was produced by these mutant plasmids. The cat ribosome-binding site, functional in E. coli, was repositioned within the pUB110 moiety and had consequently an extended homology with B. subtilis 16S rRNA, explaining the production of native enzyme.Key words: chloramphenicol acetyltransferase, Bacillus subtilis, postexponential gene expression, plasmid pUB110, ribosome-binding site, transcriptional promoter.

Binding site for fungal β-lactone hymeglusin on cytosolic 3-hydroxy-3-methylglutaryl coenzyme A synthase

2004 ◽  
Vol 1636 (1) ◽  
pp. 22-28 ◽  
Author(s):  
Hiroshi Tomoda ◽  
Naomi Ohbayashi ◽  
Yuko Morikawa ◽  
Hidetoshi Kumagai ◽  
Satoshi Ōmura
Keyword(s):  
Binding Site ◽  
Coenzyme A

scholarly journals Binding site for coenzyme A revealed in the structure of pyruvate:ferredoxin oxidoreductase from Moorella thermoacetica

2018 ◽  
Vol 115 (15) ◽  
pp. 3846-3851 ◽  
Author(s):  
Percival Yang-Ting Chen ◽  
Heather Aman ◽  
Mehmet Can ◽  
Stephen W. Ragsdale ◽  
Catherine L. Drennan
Keyword(s):  
Carbon Dioxide ◽  
Binding Site ◽  
Coenzyme A ◽  
Structural Data ◽  
Thiamine Pyrophosphate ◽  
Sequence Motifs ◽  
Moorella Thermoacetica ◽  
Microbial Enzyme ◽  
Pyruvate:Ferredoxin Oxidoreductase ◽  
Domain Iii

Pyruvate:ferredoxin oxidoreductase (PFOR) is a microbial enzyme that uses thiamine pyrophosphate (TPP), three [4Fe-4S] clusters, and coenzyme A (CoA) in the reversible oxidation of pyruvate to generate acetyl-CoA and carbon dioxide. The two electrons that are generated as a result of pyruvate decarboxylation are used in the reduction of low potential ferredoxins, which provide reducing equivalents for central metabolism, including the Wood–Ljungdahl pathway. PFOR is a member of the 2-oxoacid:ferredoxin oxidoreductase (OFOR) superfamily, which plays major roles in both microbial redox reactions and carbon dioxide fixation. Here, we present a set of crystallographic snapshots of the best-studied member of this superfamily, the PFOR from Moorella thermoacetica (MtPFOR). These snapshots include the native structure, those of lactyl-TPP and acetyl-TPP reaction intermediates, and the first of an OFOR with CoA bound. These structural data reveal the binding site of CoA as domain III, the function of which in OFORs was previously unknown, and establish sequence motifs for CoA binding in the OFOR superfamily. MtPFOR structures further show that domain III undergoes a conformational change upon CoA binding that seals off the active site and positions the thiolate of CoA directly adjacent to the TPP cofactor. These structural findings provide a molecular basis for the experimental observation that CoA binding accelerates catalysis by 105-fold.

The Conformation of Coenzyme A Bound to Chloramphenicol Acetyltransferase Determined by Transferred NOE Experiments

1996 ◽  
Vol 262 (4) ◽  
pp. 543-558 ◽  
Author(s):  
I.L. Barsukov ◽  
L.-Y. Lian ◽  
J. Ellis ◽  
K.-H. Sze ◽  
W.V. Shaw ◽  
...  
Keyword(s):  
Coenzyme A ◽  
Chloramphenicol Acetyltransferase ◽  
Transferred Noe

scholarly journals Point Mutations In Candida Glabrata 3-Hydroxy-3-Methyl Glutaryl Coenzyme A Reductase (HMGRCg) An Antifungal Target, Decrease Enzymatic Activity And Substrate/Inhibitor Affinity

2021 ◽  
Author(s):  
Dulce Andrade-Pavón ◽  
Vanessa Fernández-Muñoz ◽  
Wendy González-Ibarra ◽  
César Hernández-Rodríguez ◽  
J. Antonio Ibarra ◽  
...  
Keyword(s):  
Binding Energy ◽  
Enzymatic Activity ◽  
Binding Site ◽  
Candida Glabrata ◽  
Coenzyme A ◽  
Substrate Binding ◽  
Point Mutations ◽  
Substrate Binding Site ◽  
Substrate Inhibitor ◽  
Coenzyme A Reductase

Abstract An alternative target for antifungal drugs is 3-hydroxy-3-methyl glutaryl coenzyme A reductase (HMGR), a key enzyme in the ergosterol biosynthesis pathway. The aim of this study was to obtain, purify, characterize, and overexpress five point mutations in highly conserved regions of the catalytic domain of Candida glabrata HMGR (HMGRCg) to explore the function of key amino acid residues. Glutamic acid (Glu) was substituted by glutamine in the E680Q mutant (at the dimerization site), Glu by glutamine in E711Q (at the substrate binding site), aspartic acid by alanine in D805A and methionine by arginine in M807R (the latter two at the cofactor binding site). A double mutation, E680Q-M807R, was also made. The in vitro enzymatic activity decreased significantly in all recombinant (versus wild-type) HMGRCg, and the in silico binding energy for simvastatin, alpha-asarone and the substrate HMG-CoA was also lower for the mutants. The lowest enzymatic activity and binding energy was displayed by E711Q, suggesting that Glu711 (in the substrate binding site) is an important residue for enzymatic activity. The double mutant HMGRCg E680Q-M807R exhibited the second lowest enzymatic activity. The current findings provide insights into the role of residues in the catalytic site of HMGRCg.

Acetyl coenzyme A binding by chloramphenicol acetyltransferase: long-range electrostatic determinants of coenzyme A recognition

Biochemistry ◽  
1992 ◽  
Vol 31 (17) ◽  
pp. 4198-4205 ◽  
Author(s):  
Philip J. Day ◽  
William V. Shaw ◽  
Michael R. Gibbs ◽  
Andrew G. W. Leslie
Keyword(s):  
Long Range ◽  
Coenzyme A ◽  
Chloramphenicol Acetyltransferase ◽  
Acetyl Coenzyme A ◽  
Acetyl Coenzyme
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