ArticleProceedings of the National Academy of Sciences of the United States of America1997
Identification of active sites in amidase: evolutionary relationship between amide bond- and peptide bond-cleaving enzymes.
Article in Proceedings of the National Academy of Sciences of the United States of America, 1997. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
21 citing papers in PubMed, 113 citations in OpenAlex.
- Amidase-Catalyzed Desorption of COAngewandte Chemie (International ed. in English) · 2026Article
- New function of aldoxime dehydratase: Redox catalysis and the formation of an unexpected product.PloS one · 2017Article
- Enzymes from Extreme Environments and Their Industrial Applications.Frontiers in bioengineering and biotechnology · 2015Review
- Archaeal Enzymes and Applications in Industrial Biocatalysts.Archaea (Vancouver, B.C.) · 2015Review
- Purification and characterization of a thermostable aliphatic amidase from the hyperthermophilic archaeon Pyrococcus yayanosii CH1.Extremophiles : life under extreme conditions · 2014Article
- A new synthetic route to N-benzyl carboxamides through the reverse reaction of N-substituted formamide deformylase.Applied and environmental microbiology · 2014Article
- Strategy for successful expression of the Pseudomonas putida nitrile hydratase activator P14K in Escherichia coli.BMC biotechnology · 2013Article
- Crystal structure of aldoxime dehydratase and its catalytic mechanism involved in carbon-nitrogen triple-bond synthesis.Proceedings of the National Academy of Sciences of the United States of America · 2013Article
- Article
- Product-induced gene expression, a product-responsive reporter assay used to screen metagenomic libraries for enzyme-encoding genes.Applied and environmental microbiology · 2010Article
- Self-subunit swapping chaperone needed for the maturation of multimeric metalloenzyme nitrile hydratase by a subunit exchange mechanism also carries out the oxidation of the metal ligand cysteine residues and insertion of cobalt.The Journal of biological chemistry · 2009Article
- Discovery of posttranslational maturation by self-subunit swapping.Proceedings of the National Academy of Sciences of the United States of America · 2008Article
- Transcriptional regulation of the nitrile hydratase gene cluster in Pseudomonas chlororaphis B23.Journal of bacteriology · 2008Article
- Indole-3-acetic acid (IAA) biosynthesis in the smut fungus Ustilago maydis and its relevance for increased IAA levels in infected tissue and host tumour formation.Molecular plant pathology · 2008Article
- Discovery of a reaction intermediate of aliphatic aldoxime dehydratase involving heme as an active center.Proceedings of the National Academy of Sciences of the United States of America · 2006Article
- Amine-synthesizing enzyme N-substituted formamide deformylase: screening, purification, characterization, and gene cloning.Proceedings of the National Academy of Sciences of the United States of America · 2004Article
- Cloning and heterologous expression of an enantioselective amidase from Rhodococcus erythropolis strain MP50.Applied and environmental microbiology · 2002Article
- Complete nucleotide sequence and organization of the atrazine catabolic plasmid pADP-1 from Pseudomonas sp. strain ADP.Journal of bacteriology · 2001Article
- Identification of active-site residues in Bradyrhizobium japonicum malonamidase E2.The Biochemical journal · 2000Article
- Toc64, a new component of the protein translocon of chloroplasts.The Journal of cell biology · 2000Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Mainly based on various inhibitor studies previously performed, amidases came to be regarded as sulfhydryl enzymes. Not completely satisfied with this generally accepted interpretation, we performed a series of site-directed mutagenesis studies on one particular amidase of Rhodococcus rhodochrous J1 that was involved in its nitrile metabolism. For these experiments, the recombinant amidase was produced as the inclusion body in Escherichia coli to greatly facilitate its recovery and subsequent purification. With regard to the presumptive active site residue Cys203, a Cys203 --> Ala mutant enzyme still retained 11.5% of the original specific activity. In sharp contrast, substitutions in certain other positions in the neighborhood of Cys203 had a far more dramatic effect on the amidase. Glutamic acid substitution of Asp191 reduced the specific activity of the mutant enzyme to 1.33% of the wild-type activity. Furthermore, Asp191 --> Asn substitution as well as Ser195 --> Ala substitution completely abolished the specific activity. It would thus appear that, among various conserved residues residing within the so-called signature sequence common to all amidases, the real active site residues are Asp191 and Ser195 rather than Cys203. Inasmuch as an amide bond (CO-NH2) in the amide substrate is not too far structurally removed from a peptide bond (CO-NH-), the signature sequences of various amidases were compared with the active site sequences of various types of proteases. It was found that aspartic acid and serine residues corresponding to Asp191 and Ser195 of the Rhodococcus amidase are present within the active site sequences of aspartic proteinases, thus suggesting the evolutionary relationship between the two.
Indexed as
Identifiers
What OpenQuestion holds
Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.