Characterization of the first beta-class carbonic anhydrase from an arthropod (Drosophila melanogaster) and phylogenetic analysis of beta-class carbonic anhydrases in invertebrates

TamPub

Kuvailutiedot

dc.contributor.author Innocenti, Alessio -
dc.contributor.author Scozzafava, Andrea -
dc.contributor.author Gorr, Thomas A -
dc.contributor.author Syrjänen, Leo (Tay) -
dc.contributor.author Tolvanen, Martti (Tay) -
dc.contributor.author Hilvo, Mika (Tay) -
dc.contributor.author Olatubosun, Ayodeji (Tay) -
dc.contributor.author Leppiniemi, Jenni (Tay) -
dc.contributor.author Niederhauser, Barbara (Tay) -
dc.contributor.author Hytönen, Vesa P (Tay) -
dc.contributor.author Parkkila, Seppo (Tay) -
dc.date.accessioned 2012-06-17T20:16:26Z
dc.date.available 2012-06-16 12:07:41 -
dc.date.available 2012-06-17T20:16:26Z
dc.date.issued 2010 -
dc.identifier.issn 1471-2091 -
dc.identifier.uri http://tampub.uta.fi/handle/10024/66249
dc.description BioMed Central Open access -
dc.description.abstract Background The β-carbonic anhydrase (CA, EC 4.2.1.1) enzymes have been reported in a variety of organisms, but their existence in animals has been unclear. The purpose of the present study was to perform extensive sequence analysis to show that the β-CAs are present in invertebrates and to clone and characterize a member of this enzyme family from a representative model organism of the animal kingdom, e.g., Drosophila melanogaster. Results The novel β-CA gene, here named DmBCA, was identified from FlyBase, and its orthologs were searched and reconstructed from sequence databases, confirming the presence of β-CA sequences in 55 metazoan species. The corresponding recombinant enzyme was produced in Sf9 insect cells, purified, kinetically characterized, and its inhibition was investigated with a series of simple, inorganic anions. Holoenzyme molecular mass was defined by dynamic light scattering analysis and gel filtration, and the results suggested that the holoenzyme is a dimer. Double immunostaining confirmed predictions based on sequence analysis and localized DmBCA protein to mitochondria. The enzyme showed high CO2 hydratase activity, with a kcat of 9.5 × 105 s-1 and a kcat/KM of 1.1 × 108 M-1s-1. DmBCA was appreciably inhibited by the clinically-used sulfonamide acetazolamide, with an inhibition constant of 49 nM. It was moderately inhibited by halides, pseudohalides, hydrogen sulfide, bisulfite and sulfate (KI values of 0.67 - 1.36 mM) and more potently by sulfamide (KI of 0.15 mM). Bicarbonate, nitrate, nitrite and phenylarsonic/boronic acids were much weaker inhibitors (KIs of 26.9 - 43.7 mM). Conclusions The Drosophila β-CA represents a highly active mitochondrial enzyme that is a potential model enzyme for anti-parasitic drug development. -
dc.language.iso en -
dc.title Characterization of the first beta-class carbonic anhydrase from an arthropod (Drosophila melanogaster) and phylogenetic analysis of beta-class carbonic anhydrases in invertebrates -
dc.type fi=Artikkeli aikakauslehdessä | en=Journal article| -
dc.identifier.urn urn:nbn:uta-3-513 -
dc.identifier.doi 10.1186/1471-2091-11-28 -
dc.type.version fi=Kustantajan versio | en=Publisher's version| -
dc.subject.okm fi=Lääketieteen bioteknologia | en=Medical biotechnology| -
dc.journal.title BMC Biochemistry -
dc.journal.volume 11 -
dc.journal.number 28 -
dc.journal.volumepagerange 1-13 -
dc.oldstats 142 -

Viite kuuluu kokoelmiin:

Kuvailutiedot