The phosphocarrier protein HPr of the bacterial phosphotransferase system globally regulates energy metabolism by directly interacting with multiple enzymes in Escherichia coli

Abstract

The histidine-phosphorylatable phosphocarrier protein (HPr) is an essential component of the sugar-transporting phosphotransferase system (PTS) in many bacteria. Recent interactome findings suggested that HPr interacts with several carbohydrate-metabolizing enzymes, but whether HPr plays a regulatory role was unclear. Here, we provide evidence that HPr interacts with a large number of proteins in We demonstrate HPr-dependent allosteric regulation of the activities of pyruvate kinase (PykF, but not PykA), phosphofructokinase (PfkB, but not PfkA), glucosamine-6-phosphate deaminase (NagB), and adenylate kinase (Adk). HPr is either phosphorylated on a histidyl residue (HPr-P) or non-phosphorylated (HPr). PykF is activated only by non-phosphorylated HPr, which decreases the PykF for phosphoenolpyruvate by 10-fold (from 3.5 to 0.36 mm), thus influencing glycolysis. PfkB activation by HPr, but not by HPr-P, resulted from a decrease in the for fructose-6-P, which likely influences both gluconeogenesis and glycolysis. Moreover, NagB activation by HPr was important for the utilization of amino sugars, and allosteric inhibition of Adk activity by HPr-P, but not by HPr, allows HPr to regulate the cellular energy charge coordinately with glycolysis. These observations suggest that HPr serves as a directly interacting global regulator of carbon and energy metabolism and probably of other physiological processes in enteric bacteria.

Global landscape of cell envelope protein complexes in Escherichia coli

“…While the few remaining (<1%, 104) prey–prey associations were among cytoplasmic protein pairs, we also gained meaningful functional insights from these subnetworks into membrane transport metabolons (i.e., physical coupling of transporters and enzymes; see below) and other pathways 22 (Fig. 1e and Supplementary Note 1).…”

“…For instance, fructose PTS permease (FruA) co-purified with enzyme IIC constituents (e.g., GatC, NagE, TreB, MngA) in addition to its cognate energy-coupling factor, FruB 33 . Although not strictly membrane-bound 22 , HPr also co-purified with multiple regulatory targets (e.g., Adk, DhaK, PfkB, PykF; Supplementary Note 2). HPr is known to bind pyruvate kinase A (PykA) in the opportunistic pathogen Vibrio vulnificus 34 , whereas E. coli HPr interacts with an isozyme, pyruvate kinase 1 (PykF).…”

Cajanin Stilbene Acid Inhibited Vancomycin-Resistant Enterococcus by Inhibiting Phosphotransferase System

“…PTS system are widespread in bacteria, playing important roles in a catalytic carbohydrate concomitant uptake and phosphorylation, it is also involved in carbohydrate transport, signal transduction, and catabolite repression (Opsata et al, 2010). PTS system ensures the efficient utilization of carbohydrate bacteria in a complex environment, and the steady growth and reproduction of bacteria (Rodionova et al,2017). Previous reports suggested that PTS may be an important drug target (Huang et al, 2013).…”

UDP-glucose dehydrogenase Ugd inE. coliis activated by Gmd and RffD, is inhibited by CheY, and regulates swarming

“…Purified WecC, CheY, RssB, FolC and RnhB proteins, involved in Ugd interactions ( Table 2) (14), were tested for regulation of Ugd activity in the presence and absence of ATP. Phosphofructokinase (PfkB), previously characterised (21), was used as a negative control.…”

“…The calculated Kd for the WecC-Ugd interaction was calculated to be 0.24±0.01 µM of WecC. Phosphofructokinase PfkB (21) was used as a negative control, and no activation of Ugd was observed when this protein was added at a concentration of 0.6 µM (Fig.3).…”