- Title
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Genetic basis of thiaminase I activity in a vertebrate, zebrafish Danio rerio
- Authors
- Richter, C.A., Evans, A.N., Heppell, S.A., Zajicek, J.L., Tillitt, D.E.
- Source
- Full text @ Sci. Rep.
Identification of putative zebrafish (Danio rerio) thiaminases. Sequence alignment of previously reported thiaminase I peptide fragments from red cornetfish (Fistularia petimba) with candidate zebrafish thiaminase I protein sequences. (A) N-terminal thiaminase I peptide fragment from red cornetfish aligned with residues 25–40 of zebrafish protein NP_001314821 and residues 39–56 of zebrafish protein NP_001314926. (B) Internal thiaminase I peptide fragment from red cornetfish aligned with residues 195–204 of zebrafish protein \erm_text":"NP_001314821"}}NP_001314821 and residues 210–219 of zebrafish protein NP_001314926. |
Alignment of known and putative thiaminases. Protein sequence alignment of candidate zebrafish (Danio rerio) tenA-like thiaminase I protein sequences with candidate common carp (Cyprinus carpio) tenA-like thiaminase I ({"type":"entrez-protein","attrs":{"text":"XP_042594753","term_id":"2073630299","term_text":"XP_042594753"}}XP_042594753), candidate alewife (Alosa pseudoharengus) tenA-like thiaminase I, and thiaminase II protein TenA (Bacillus subtilis, NP_389047). The position of the TenA active site cysteine is marked with a red box. |
The zebrafish (Danio rerio) tenA-like thiaminase I gene encodes a functional thiaminase. Solubility and thiaminase I activity of recombinant candidate thiaminase I genes overexpressed in Escherichia coli, stained for activity showing thiamine degradation as clear areas, and with subsequent Coomassie blue protein stain. Gels are cropped to show relevant lanes and mobility range; original gels are presented in Supplementary Fig. 9. Panel A, insoluble fractions; Panel B, soluble fractions; lane 1, common carp (Cyprinus carpio), lane 2, zebrafish; lane 3, alewife (Alosa pseudoharengus); lane 4, Paenibacillus thiaminolyticus; lane 5, empty pET52b vector. |