Preliminary characterization of two novel thiocyanate dehydrogenases from halophilic sulfur oxidizing Gammaproteobacteria Thiohalobacter thiocyanaticus HRh1T and Guyparkeria sp. SCN-R1

Shipkov N.S., Dergousova N.I., Rakitina T.V., Tsallagov S.I., Tikhonova T.V., Popov V.O., Dimitry Y. Sorokin

Research output: Contribution to conferenceAbstractScientific

Abstract

CopC is a small periplasmic protein from haloalkaliphilic bacteria of the genus Thioalkalivibrio. This CopC differs from other well-known CopC proteins by the presence only
Cu(II) binding site. By tryptophan fluorescence quenching method we showed that CopC can bind only one Cu(II) ion. opC belongs to the high-affinity CopC subfamily (С0-2) with conserved N-terminal His1- Xxx-His3 motif [Lawton et al., 2016]. It is known that CopC proteins can play the role of copper chaperon [Hussain et al., 2007]. In this regard Thiocyanate dehydrogenase (TcDH) as a copper containing protein from the same bacteria was chosen. When the bacterium Tv. paradoxus ARh1
grows with thiocyanate, the expression level of CopCD genes increases in parallel with that of TcDH genes. Genes of copper-binding proteins CopCD are located before the TcDH gene in Thioalkalivibrio genomes. TcDH apo form (containing no copper ions) can be activated by the Cu(II) sarurated CopC
more effective than that of Cu(II) ions at the same concentration. By tryptophan fluorescence quenching assay we revealed the complex formation (TcDH and
CopC) with stoichiometric ratio 1:2. After complex separating using gel-filtration the increasing of copper content in the molecule of TcDH with decreasing of that in the molecule of CopC was observed. The activity of copper-rich TcDH is also increased in comparison with TcDH apo form. The data obtained indicate the possibility of TcDH to form a complex with CopC with the subsequent transfer of copper ions to the TcDH molecule. These data confirm that CopC can play the role of chaperone responsible for incorporation of copper into the TcDH active site.
Original languageEnglish
Pages117
Publication statusPublished - 2019
Event12th International Conference BIOCATALYSIS-2019: Fundamentals & Applications - St.Petersburg, Russian Federation
Duration: 24 Jun 201928 Jun 2019

Conference

Conference12th International Conference BIOCATALYSIS-2019: Fundamentals & Applications
CountryRussian Federation
CitySt.Petersburg
Period24/06/1928/06/19

Fingerprint Dive into the research topics of 'Preliminary characterization of two novel thiocyanate dehydrogenases from halophilic sulfur oxidizing Gammaproteobacteria Thiohalobacter thiocyanaticus HRh1T and Guyparkeria sp. SCN-R1'. Together they form a unique fingerprint.

Cite this