Isolation of recombinant cysteine dioxygenase protein from Trichophyton mentagrophytes

0368595 - ÚEB 2012 RIV DE eng J - Journal Article
Kašperová, A. - Kunert, J. - Horynová, M. - Weigl, E. - Sebela, M. - Lenobel, René - Raška, M.
Isolation of recombinant cysteine dioxygenase protein from Trichophyton mentagrophytes.
Mycoses. Roč. 54, č. 5 (2011), E456-E462. ISSN 0933-7407. E-ISSN 1439-0507
R&D Projects: GA ČR GA301/08/1649
Institutional research plan: CEZ:AV0Z50380511
Keywords : Cysteine dioxygenase * dermatophytes * recombinant protein * keratinolytic fungi * cDNA
Subject RIV: CE - Biochemistry
Impact factor: 2.247, year: 2011

Cysteine dioxygenase (CDO, EC 1.13.11.20) catalyses the oxygenation of cysteine to cysteine sulphinic acid leading to the production of sulphite, sulphate and taurine as the final metabolites of cysteine catabolism. Keratinolytic fungi secrete sulphite and sulphate to reduce disulphide bridges in host tissue keratin proteins as the first step of keratinolysis. In the present study, we describe the identification of cDNA, as well as expression and characterisation of recombinant CDO protein from Trichophyton mentagrophytes. The cDNA was amplified using primers designed on the basis of high conservancy CDO regions identified in other fungi. PCR product was cloned and sequenced. Recombinant CDO was expressed in Escherichia colt, and affinity purified and identified by matrix-assisted laser desorption/ionization - time-of-flight mass spectrometry (MALDI-TOF MS). Enzyme activity was assayed by monitoring the production of cysteine sulphinate using mass spectrometry.
Permanent Link: http://hdl.handle.net/11104/0202894