Control of AmtB-GlnK complex formation by intracellular levels of ATP, ADP, and 2-oxoglutarate.
P(II) proteins are one of the most widespread families of signal transduction proteins in nature, being ubiquitous throughout bacteria, archaea, and plants. They play a major role in coordinating nitrogen metabolism by interacting with, and regulating the activities of, a variety of enzymes, transcription factors, and membrane transport proteins. The regulatory properties of P(II) proteins derive from their ability to bind three effectors: ATP, ADP, and 2-oxoglutarate. However, a clear model to integrate physiological changes with the consequential structural changes that mediate P(II) interaction with a target protein has so far not been developed. In this study, we analyzed the fluctuations in intracellular effector pools in Escherichia coli during association and dissociation of the P(II) protein GlnK with the ammonia channel AmtB. We determined that key features promoting AmtB-GlnK complex formation are the rapid drop in the 2-oxoglutarate pool upon ammonium influx and a simultaneous, but transient, change in the ATP/ADP ratio. We were also able to replicate AmtB-GlnK interactions in vitro using the same effector combinations that we observed in vivo. This comprehensive data set allows us to propose a model that explains the way in which interactions between GlnK and its effectors influence the conformation of GlnK and thereby regulate its interaction with AmtB.
Radchenko
Martha V
MV
Department of Molecular Microbiology, John Innes Centre, Norwich NR4 7UH, United Kingdom.
Thornton
Jeremy
J
Merrick
Mike
M
D000244:Adenosine Diphosphate / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism*; D027682:Cation Transport Proteins / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000378:metabolism*; D017136:Ion Transport / Q000502:physiology; D007656:Ketoglutaric Acids / Q000378:metabolism*; D008954:Models, Biological; D046912:Multiprotein Complexes / Q000378:metabolism*; D009713:Nucleotidyltransferases / Q000378:metabolism*; D051922:PII Nitrogen Regulatory Proteins / Q000378:metabolism*; D000644:Quaternary Ammonium Compounds / Q000378:metabolism
C111096:AmtB protein, E coli; D027682:Cation Transport Proteins; D029968:Escherichia coli Proteins; D007656:Ketoglutaric Acids; D046912:Multiprotein Complexes; D051922:PII Nitrogen Regulatory Proteins; D000644:Quaternary Ammonium Compounds; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; C497878:glnK protein, E coli; D009713:Nucleotidyltransferases