PMID:17644595

Citation	Wolfe, DM, Zhang, Y and Roberts, GP (2007) Specificity and regulation of interaction between the PII and AmtB1 proteins in Rhodospirillum rubrum. J. Bacteriol. 189:6861-9
Abstract	The nitrogen regulatory protein P(II) and the ammonia gas channel AmtB are both found in most prokaryotes. Interaction between these two proteins has been observed in several organisms and may regulate the activities of both proteins. The regulation of their interaction is only partially understood, and we show that in Rhodospirillum rubrum one P(II) homolog, GlnJ, has higher affinity for an AmtB(1)-containing membrane than the other two P(II) homologs, GlnB and GlnK. This interaction strongly favors the nonuridylylated form of GlnJ and is disrupted by high levels of 2-ketoglutarate (2-KG) in the absence of ATP or low levels of 2-KG in the presence of ATP. ADP inhibits the destabilization of the GlnJ-AmtB(1) complex in the presence of ATP and 2-KG, supporting a role for P(II) as an energy sensor measuring the ratio of ATP to ADP. In the presence of saturating levels of ATP, the estimated K(d) of 2-KG for GlnJ bound to AmtB(1) is 340 microM, which is higher than that required for uridylylation of GlnJ in vitro, about 5 microM. This supports a model where multiple 2-KG and ATP molecules must bind a P(II) trimer to stimulate release of P(II) from AmtB(1), in contrast to the lower 2-KG requirement for productive uridylylation of P(II) by GlnD.
Links	PubMed PMC2045211 Online version:10.1128/JB.00759-07
Keywords	Adenosine Diphosphate/pharmacology; Adenosine Triphosphate/pharmacology; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Blotting, Western; Cation Transport Proteins/genetics; Cation Transport Proteins/metabolism; Ketoglutaric Acids/pharmacology; PII Nitrogen Regulatory Proteins/genetics; PII Nitrogen Regulatory Proteins/metabolism; Protein Binding/drug effects; Quaternary Ammonium Compounds/metabolism; Rhodospirillum rubrum/genetics; Rhodospirillum rubrum/metabolism
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
RHORT:GLNB		GO:0006808: regulation of nitrogen utilization	ECO:0000314:		P		Figure 4. Quantitative analysis of PII rebinding to the amtBI	complete CACAO 3297

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