title: Protonography, a technique applicable for the analysis of η-carbonic anhydrase activity.
abstract: Protonography, a sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) technique derived from zymography was recently reported by our group to be an effective, cheap and reproducible technique for evidencing catalytically active α-carbonic anhydrase (CA, EC 4.2.1.1) isoforms, such as the bovine red blood cell isoform bCA or the bacterial enzyme from Vibrio cholerae, VchCA. CA activity was also observed on the protonogram of a cellular extract of Escherichia coli, evidencing the presence of one or more β-class such enzymes. Here we show that protonography can also be applied to the recently discovered η-CA family using the Plasmodium falciparum enzyme PfCA as an example. The protonogram of PfCA clearly showed catalytically active η-CA with a specific band at 22.0 kDa, which was quite distinct from the band of the red blood cell bovine enzyme bCA, which was observed at 28.8 kDa. The different migration pattern of α- and η-CAs might be a useful tool to detect Plasmodium falciparum in infected human red blood cells by an easy, routine inexpensive technique.
authors: a CNR - Istituto di Bioscienze e Biorisorse , Napoli , Italy and.|b Dipartimento Neurofarba, Sezione di Scienze Farmaceutiche, and Laboratorio di Chimica Bioinorganica, Polo Scientifico , Università Degli Studi di Firenze , Florence , Italy.
authors: Sonia
authors: S
authors: Del Prete
authors: a CNR - Istituto di Bioscienze e Biorisorse , Napoli , Italy and.
authors: Viviana
authors: V
authors: De Luca
authors: b Dipartimento Neurofarba, Sezione di Scienze Farmaceutiche, and Laboratorio di Chimica Bioinorganica, Polo Scientifico , Università Degli Studi di Firenze , Florence , Italy.
authors: Claudiu T
authors: CT
authors: Supuran
authors: a CNR - Istituto di Bioscienze e Biorisorse , Napoli , Italy and.
authors: Clemente
authors: C
authors: Capasso
mesh_terms: D000818:Animals; D055162:Biocatalysis; D002256:Carbonic Anhydrases / Q000235:genetics / Q000378:metabolism*; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D004789:Enzyme Activation; D057075:Enzyme Assays / Q000379:methods*; D004912:Erythrocytes / Q000201:enzymology; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D010963:Plasmodium falciparum / Q000201:enzymology; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry*; D014734:Vibrio cholerae / Q000201:enzymology
chemical_list: D007527:Isoenzymes; D012967:Sodium Dodecyl Sulfate; D002256:Carbonic Anhydrases
keywords: carbonic anhydrase; hydratase activity; metalloenzymes; protonography; η-Class enzyme