TY - JOUR
T1 - C4-dicarboxylate metabolons
T2 - interaction of C4-dicarboxylate transporters of Escherichia coli with cytosolic enzymes
AU - Schubert, Christopher
AU - Kim, Nam Yeun
AU - Unden, Gottfried
AU - Kim, Ok Bin
N1 - Funding Information:
Acknowledgments We are grateful to Dr A. Strecker (Mainz) for preparing the plasmids with the DcuASWT25 and DcuCSWT25 constructs. Financial support from the Deutsche Forschungsgemeinschaft (grant UN 49/19-1) and from the National Research Foundation of Korea (NRF-2019R1A2C1008066) is gratefully acknowledged
Publisher Copyright:
© 2022 The Author(s). Published by Oxford University Press on behalf of FEMS.
PY - 2022
Y1 - 2022
N2 - Metabolons represent the structural organization of proteins for metabolic or regulatory pathways. Here, the interaction of fumarase FumB, aspartase AspA, and L-tartrate dehydratase TtdAB with the C4-dicarboxylate (C4-DC) transporters DcuA, DcuB, DcuC, and the L-tartrate transporter TtdT of Escherichia coli was tested by a bacterial two-hybrid (BACTH) assay in situ, or by co-chromatography using mSPINE (membrane Streptavidin protein interaction experiment). From the general C4-DC transporters, DcuB interacted with FumB and AspA, DcuA with AspA, whereas DcuC interacted with neither FumB nor AspA. Moreover, TtdT did not interact with TtdAB. The fumB-dcuB, the dcuA-aspA, and the ttdAB-ttdT genes encoding the respective proteins colocalize on the genome and each pair of genes forms cotranscripts, whereas the dcuC gene lies alone. The data suggest the formation of DcuB/FumB and DcuB/AspA metabolons for the uptake of L-malate, or L-aspartate, and their conversion to fumarate for fumarate respiration and excretion of the product succinate. The DcuA/AspA metabolon catalyzes uptake and conversion of L-aspartate to fumarate coupled to succinate excretion. The DcuA/AspA metabolon provides ammonia at the same time for nitrogen assimilation (ammonia shuttle). On the other hand, TtdT and TtdAB are not organized in a metabolon. Reasons for the formation (DcuA/AspA, DcuB/FumB, and DcuB/AspA) or nonformation (DcuC, TtdT, and TtdAB) of metabolons are discussed based on their metabolic roles.
AB - Metabolons represent the structural organization of proteins for metabolic or regulatory pathways. Here, the interaction of fumarase FumB, aspartase AspA, and L-tartrate dehydratase TtdAB with the C4-dicarboxylate (C4-DC) transporters DcuA, DcuB, DcuC, and the L-tartrate transporter TtdT of Escherichia coli was tested by a bacterial two-hybrid (BACTH) assay in situ, or by co-chromatography using mSPINE (membrane Streptavidin protein interaction experiment). From the general C4-DC transporters, DcuB interacted with FumB and AspA, DcuA with AspA, whereas DcuC interacted with neither FumB nor AspA. Moreover, TtdT did not interact with TtdAB. The fumB-dcuB, the dcuA-aspA, and the ttdAB-ttdT genes encoding the respective proteins colocalize on the genome and each pair of genes forms cotranscripts, whereas the dcuC gene lies alone. The data suggest the formation of DcuB/FumB and DcuB/AspA metabolons for the uptake of L-malate, or L-aspartate, and their conversion to fumarate for fumarate respiration and excretion of the product succinate. The DcuA/AspA metabolon catalyzes uptake and conversion of L-aspartate to fumarate coupled to succinate excretion. The DcuA/AspA metabolon provides ammonia at the same time for nitrogen assimilation (ammonia shuttle). On the other hand, TtdT and TtdAB are not organized in a metabolon. Reasons for the formation (DcuA/AspA, DcuB/FumB, and DcuB/AspA) or nonformation (DcuC, TtdT, and TtdAB) of metabolons are discussed based on their metabolic roles.
KW - BACTH
KW - C-dicarboxylate transporter
KW - Escherichia coli
KW - L-aspartate
KW - mSPINE
KW - metabolon
UR - http://www.scopus.com/inward/record.url?scp=85138491962&partnerID=8YFLogxK
U2 - 10.1093/femsle/fnac078
DO - 10.1093/femsle/fnac078
M3 - Article
C2 - 36044995
AN - SCOPUS:85138491962
SN - 0378-1097
VL - 369
JO - FEMS Microbiology Letters
JF - FEMS Microbiology Letters
IS - 1
M1 - fnac078
ER -