title: Protonography, a powerful tool for analyzing the activity and the oligomeric state of the γ-carbonic anhydrase identified in the genome of Porphyromonas gingivalis.
abstract: Carbonic anhydrases (CAs, EC 4.2.1.1) are metalloenzymes, mostly containing zinc within their active site, which catalyze a simple but physiologically relevant reaction in all life kingdoms, carbon dioxide hydration to bicarbonate and protons. Six CA classes (α, β, γ, δ, ζ and η) and multiple CA isoforms evolved in organisms all over the phylogenetic tree, for facing the need to efficiently convert high amounts of CO2 to its hydration products. These enzymes are thus involved in many physiologic processes, such as photosynthesis, respiration, CO2 transport, electrolyte secretion in many tissues/organs; biosynthetic reactions (gluconeogenesis, lipogenesis, ureagenesis), etc. Recently, our group reported a new technique to assay CA activity on SDS-PAGE gels, named 'protonography' due to its similarity to zymography. By using protonography, the conversion of CO2 into protons can be visualized as a yellow band on a polyacrylamide gel. By using this technique we demonstrated the possibility to detect activity of the α-CA from Vibrio cholerae as well as the β- and γ-CAs present in Escherichia coli extracts. Furthermore, the activity of the newly discovered η-class enzyme from Plasmodium falciparum has also been evidenced with protonography, illustrating its wide use. Here we show that protonography can be also useful to reveal the oligomeric state of the γ-CA identified in the genome of the bacterial parasite colonizing the oral cavity, Porphyromonas gingivalis, possibly allowing for a simple and efficient diagnostic method.
authors.affiliation: Istituto di Bioscienze e Biorisorse, CNR, Via Pietro Castellino 111, 80131 Napoli, Italy; Laboratorio di Chimica Bioinorganica, Polo Scientifico, Università degli Studi di Firenze, Sesto Fiorentino, Firenze, Italy.
authors.forename: Sonia
authors.initials: S
authors.lastname: Del Prete
authors.affiliation: Istituto di Bioscienze e Biorisorse, CNR, Via Pietro Castellino 111, 80131 Napoli, Italy.
authors.forename: Viviana
authors.initials: V
authors.lastname: De Luca
authors.affiliation: Istituto di Bioscienze e Biorisorse, CNR, Via Pietro Castellino 111, 80131 Napoli, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Iandolo
authors.affiliation: Laboratorio di Chimica Bioinorganica, Polo Scientifico, Università degli Studi di Firenze, Sesto Fiorentino, Firenze, Italy; Dipartimento di Scienze Farmaceutiche, Polo Scientifico, Università degli Studi di Firenze, Sesto Fiorentino, Firenze, Italy.
authors.forename: Claudiu T
authors.initials: CT
authors.lastname: Supuran
authors.affiliation: Istituto di Bioscienze e Biorisorse, CNR, Via Pietro Castellino 111, 80131 Napoli, Italy. Electronic address: clemente.capasso@ibbr.cnr.it.
authors.forename: Clemente
authors.initials: C
authors.lastname: Capasso
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D002245:Carbon Dioxide / Q000737:chemistry / Q000378:metabolism; D002256:Carbonic Anhydrases / Q000737:chemistry* / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D016680:Genome, Bacterial; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D010963:Plasmodium falciparum / Q000737:chemistry / Q000201:enzymology; D016966:Porphyromonas gingivalis / Q000737:chemistry* / Q000201:enzymology; D021122:Protein Subunits / Q000737:chemistry* / Q000235:genetics; D011522:Protons; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D014734:Vibrio cholerae / Q000737:chemistry / Q000201:enzymology
chemical_list: D001426:Bacterial Proteins; D021122:Protein Subunits; D011522:Protons; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D002245:Carbon Dioxide; D002256:Carbonic Anhydrases
keywords: Carbonic anhydrase; Hydratase activity; Metalloenzymes; Protonography