N-acetylprocainamide [BSA]

Much of the research on Na+/H+ exchange has been done in prokaryotic models, mainly on the NhaA Na+/H+-exchanger from Escherichia coli (EcNhaA). Two conserved aspartate residues, Asp-163 and Asp-164, are essential for transport and are candidates for possible binding sites for the two H+ that are exchanged for one Na+ to make the overall transport process electrogenic. More recently, a proposed mechanism of transport for EcNhaA has suggested direct binding of one of the transported H+ to the conserved Lys-300 residue, a salt bridge partner of Asp-163. This contention is supported by a study reporting that substitution of the equivalent residue, Lys-305, of a related Na+/H+ antiporter, NapA from Thermus thermophilus, renders the transporter electroneutral. In this work, we sought to establish whether the Lys-300 residue and its partner Asp-163 are essential for the electrogenicity of EcNhaA. To that end, we replaced Lys-300 with Gln, either alone or together with the simultaneous substitution of Asp-163 with Asn, and characterized these transporter variants in electrophysiological experiments combined with H+ transport measurements and stability analysis. We found that K300Q EcNhaA can still support electrogenic Na+/H+ antiport in EcNhaA, but has reduced thermal stability. A parallel electrophysiological investigation of the K305Q variant of TtNapA revealed that it is also electrogenic. Furthermore, replacement of both salt bridge partners in the ion-binding site of EcNhaA produced an electrogenic variant (D163N/K300Q). Our findings indicate that alternative mechanisms sustain EcNhaA activity in the absence of canonical ion-binding residues and that the conserved lysines confer structural stability.

Colwellia sp. Arc7-635, a psychrophilic denitrifying bacterium isolated from Arctic seawater, uses NO3- or NH4+ as the sole nitrogen source to grow at low temperatures. In this article, we describe the complete genome of Colwellia sp. Arc7-635. The genome has one circular chromosome of 4,741,350bp (38.41mol% G+C content), consisting of 3841 coding genes, 91 tRNA genes, as well as seven rRNA operons of 16S-23S-5S rRNA, and one operon of 16S-23S-5S-5S rRNA. According to the genomic annotation results, strain Colwellia sp. Arc7-635 encodes a complete denitrifying pathway consisting of genes affiliated with nitrate reductase, nitrite reductase, nitric oxide reductase, and nitrous oxide reductase. Genes affiliated with nitrate reduction to ammonia including nitrate reductases (NapA and NapB) and nitrite reductases (NirA, NirB, and NirD) were also identified. The whole genome sequences of Arc7-635 provide information that is useful for further clarifying nitrogen metabolisms and facilitate its potential applications in the bioremediation of nitrogen pollutions.