In contrast, a more recent study found that CheA could bind to the receptors independent of CheW and that CheW only strengthened the interaction [86]. An in vivo localization study found that truncated CheA constructs could bind to receptor clusters independently of CheW, whereas full-length VX-680 CheA required CheW for this [87]. Only Htr group 1 matches the expected composition of Selleck SBE-��-CD prokaryotic

taxis signaling complexes (receptor-transducer, CheW, CheA, CheY, [19, 73]). Considering that binding of a CheW domain protein is mandatory for CheA activity [88–93], our findings indicate that only the receptors from group 1 were active under the tested conditions. At least for Htr11 (Car, the cytoplasmic arginine receptor, [42]), the only receptor with known signal that was assigned to a group other than group 1, this would make sense. Hbt.salinarum degrades arginine to ornithine coupled with the production of ATP [94]. This substrate-level phosphorylation allows the cells to grow in the absence

of light and oxygen, making taxis towards arginine crucial under these conditions. Under the aerobic conditions used in our experiments, the cells can produce energy by oxidative phosphorylation. Arginine is indeed metabolized under aerobic conditions and is depleted rapidly from the medium, but it can be resynthesized from ornithine [95]. Consequently, the cells have no need for arginine uptake and arginine taxis could be switched off. Two novel interactors WH-4-023 manufacturer of CheA Two proteins were identified as novel interaction partners of CheA (Figures 3 and 5). The first is PurNH (OE1620R) which is annotated as a phosphoribosylglycinamide formyltransferase (EC 2.1.2.2) / phosphoribosylaminoimidazolecarboxamide formyltransferase (EC 2.1.2.3). Thus it carries out two essential enzymatic activities in purine metabolism. PurNH was fished by CheA, CheW1 and CheY (Figure 5).

When PurNH was subsequently used as bait, it fished CheA and most of the group 1 Htrs. In all experiments, PurNH showed an interaction and exchange behavior identical to that of CheA (Additional file 4), indicating that it is statically bound to CheA. Figure 5 Interactions of the core signaling proteins CheW1 and CheA and their novel interaction partners PurNH and OE4643R. Plots show Grape seed extract the association score of the proteins identified in one-step (A-D) or two-step (E-H) bait fishing experiments with CheW1 (A, E), CheA (B, F), PurNH (C, G) and OE4643R (D, H). The dashed line indicates the threshold used in this study for assuming an interaction. The proteins CheA, CheW1, CheW2, PurNH and OE4643R are labeled in the plots when identified with an association score above the threshold. For the underlying data see Additional file 3 and Additional file 4. The second novel interactor is OE4643R, a conserved protein of unknown function. OE4643R belongs to the uncharacterized protein family DUF151 (Pfam, [96]) and the cluster of orthologous groups COG1259 (“uncharacterized conserved protein”) [97, 98].