Open AccessCorrinoid activation by a RACE protein: studies on the interaction of the proteins involved
Author(s) -
Nguyen Hai Dang,
Studenik Sandra,
Diekert Gabriele
Publication year - 2013
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/1574-6968.12178
Subject(s) - corrinoid , methyltransferase , demethylase , biochemistry , enzyme , western blot , chemistry , demethylation , substrate (aquarium) , stereochemistry , biology , methylation , gene expression , gene , epigenetics , ecology , dna methylation
The O ‐demethylases of anaerobes are corrinoid‐dependent, ether‐cleaving methyltransferase enzyme systems consisting of four components. The interaction of the O ‐demethylase components of the acetogenic bacterium Acetobacterium dehalogenans was studied by protein mobility on native PAGE , far‐Western blot analysis and yeast two‐hybrid screen. Using native PAGE and far‐Western blot, the interaction of the activating enzyme (AE) with its substrate, the corrinoid protein (CP), could be observed. The interaction occurred with four different CPs of A. dehalogenans and a CP from Desulfitobacterium hafniense DCB ‐2, all involved in ether cleavage. In the corrinoid reduction assay, the AE reduced all CPs tested. This result indicates a broad substrate specificity of the AE of A. dehalogenans . In addition, an interaction of the A. dehalogenans CP of the vanillate‐ O ‐demethylase with the two methyltransferases of the same enzyme system was observed. The interaction of the ether‐cleaving methyltransferase with the CP appeared to be significantly less pronounced than that reported for the homologous methanol and methylamine methyltransferase systems of methanogenic archaea.