Open AccessThe phosphotransferase system of Streptomyces coelicolor
Author(s) -
Kamionka Annette,
Parche Stephan,
Nothaft Harald,
Siepelmeyer Jörg,
Jahreis Knut,
Titgemeyer Fritz
Publication year - 2002
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1046/j.1432-1033.2002.02864.x
Subject(s) - pep group translocation , streptomyces coelicolor , catabolite repression , biochemistry , phosphotransferase , complementation , escherichia coli , biology , chemistry , microbiology and biotechnology , phosphorylation , gene , mutant
We have investigated the crr gene of Streptomyces coelicolor that encodes a homologue of enzyme IIA Glucose of Escherichia coli , which, as a component of the phospho enol pyruvate‐dependent sugar phosphotransferase system (PTS) plays a key role in carbon regulation by triggering glucose transport, carbon catabolite repression, and inducer exclusion. As in E. coli , the crr gene of S. coelicolor is genetically associated with the ptsI gene that encodes the general phosphotransferase enzyme I. The gene product IIA Crr was overproduced, purified, and polyclonal antibodies were obtained. Western blot analysis revealed that IIA Crr is expressed in vivo . The functionality of IIA Crr was demonstrated by phospho enol pyruvate‐dependent phosphorylation via enzyme I and the histidine‐containing phosphoryl carrier protein HPr. Phosphorylation was abolished when His72, which corresponds to the catalytic histidine of E. coli IIA Glucose , was mutated. The capacity of IIA Crr to operate in sugar transport was shown by complementation of the E. coli glucose‐PTS. The striking functional resemblance between IIA Crr and IIA Glucose was further demonstrated by its ability to confer inducer exclusion of maltose to E. coli . A specific interaction of IIA Crr with the maltose permease subunit MalK from Salmonella typhimurium was uncovered by surface plasmon resonance. These data suggest that this IIA Glucose ‐like protein may be involved in carbon metabolism in S. coelicolor .