Recombinant Nostoc sp. Potassium-transporting ATPase C chain 2 (kdpC2)
Description
Introduction to Recombinant Nostoc sp. Potassium-transporting ATPase C chain 2 (kdpC2)
Recombinant Nostoc sp. Potassium-transporting ATPase C chain 2 (kdpC2) refers to a genetically engineered form of the KdpC subunit derived from Nostoc species, which is a genus of cyanobacteria known for its ability to fix nitrogen and form filamentous colonies . The KdpC subunit is part of the Kdp system, a high-affinity potassium uptake system found in bacteria and archaea . This system is crucial for maintaining potassium homeostasis, which is essential for various cellular processes, including enzyme activity, protein synthesis, and regulation of cell volume .
Structure and Function of Kdp System
The Kdp system is an ATP-dependent potassium transporter composed of three main subunits: KdpA, KdpB, and KdpC .
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KdpA: This subunit is the potassium-selective pore through which potassium ions are transported across the cell membrane.
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KdpB: This subunit is an ATPase that provides the energy for potassium transport by hydrolyzing ATP.
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KdpC: This subunit is a small, membrane-associated protein that modulates the activity of the KdpAB complex.
The KdpC subunit is essential for the proper function and regulation of the Kdp system . It interacts with the KdpAB complex, influencing its activity and potassium affinity. The precise mechanism by which KdpC modulates KdpAB activity is still under investigation, but it is believed to involve conformational changes within the complex .
Genetic and Evolutionary Aspects
The genes encoding the Kdp system are highly conserved in bacteria and archaea, indicating their importance for survival. Phylogenetic analyses suggest that the Kdp system has evolved through horizontal gene transfer, allowing different species to acquire this essential potassium uptake system. Studies involving ancestral gene reconstruction have shown how mutations in proteins can fine-tune their functions . Key mutations can eliminate certain responses, leading to highly specific receptors, and stabilizing permissive mutations are essential for the function-switching mutations to be tolerated .
Role of Nostoc in Nitrogen Fixation and Heterocyst Formation
Nostoc species are unique because they can differentiate specialized cells called heterocysts, which are responsible for nitrogen fixation under anaerobic conditions . Heterocysts develop in response to nitrogen deprivation and are critical for the survival of Nostoc in nitrogen-limited environments . The differentiation and function of heterocysts are tightly regulated, involving various metabolic pathways and regulatory proteins .
Proteomic Studies of Nostoc
Proteomic studies have provided valuable insights into the molecular mechanisms underlying various processes in Nostoc, including the response to environmental stress and the regulation of nitrogen and carbon metabolism . For example, studies have examined the effects of β-methylamino-L-alanine (BMAA) on Nostoc under nitrogen starvation conditions, revealing that BMAA disrupts proteins involved in nitrogen and carbon metabolic pathways .
Nostocyclopeptides and Their Biological Activities
Nostoc species also produce a variety of bioactive secondary metabolites, including nostocyclopeptides (Ncps) . These nonribosomal peptides have been shown to inhibit organic anion transporters and prevent the transport of toxic microcystins and nodularin into hepatocytes . Genetic analysis of Nostoc sp. has revealed the presence of Ncp gene clusters responsible for the synthesis of these peptides .
Biotechnological Potential of Nostoc
Nostoc has several biotechnological applications, including its use as a biofertilizer, a source of bioactive compounds, and a potential platform for bioremediation . Its ability to fix nitrogen, produce exopolysaccharides, and synthesize valuable secondary metabolites makes it a promising candidate for sustainable agriculture and environmental applications .
Growth and Cultivation of Nostoc
The cultivation of Nostoc under controlled laboratory conditions is essential for research and biotechnological applications . Factors such as light intensity, nutrient availability, and medium composition can significantly affect the growth rate and biomass yield of Nostoc . Studies have shown that supplementation with nitrogen can increase the growth rate of Nostoc, while other supplements, such as magnesium, may have inhibitory effects .
Product Specs
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Target Background
KEGG: ana:all4243
STRING: 103690.all4243
Q&A
Basic Research Questions
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What is Recombinant Nostoc sp. Potassium-transporting ATPase C chain 2 (kdpC2) and what are its key characteristics?
Recombinant Nostoc sp. Potassium-transporting ATPase C chain 2 (kdpC2) is a protein component of the high-affinity potassium uptake system in the cyanobacterium Nostoc sp. It functions as part of the KdpFABC complex, which is essential for potassium homeostasis particularly under low potassium conditions. The recombinant protein is typically expressed in E. coli expression systems for research purposes .
Key Characteristics:
Property Information UniProt Accession Q8YPF1 Protein Length 199 amino acids (full length) Molecular Function Component of potassium transport system Ordered Locus Name all4243 Recommended Storage -20°C to -80°C Working Storage 4°C for up to one week Buffer Composition Tris-based buffer with 50% glycerol The protein is typically provided as a recombinant product with a His-tag or other affinity tags to facilitate purification and downstream applications .
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How does kdpC2 function within the KdpFABC complex?
The kdpC2 protein functions as a catalytic chaperone within the KdpFABC complex, playing a crucial role in the nucleotide-binding mechanism. Unlike typical P-type ATPases, the KdpFABC complex represents a unique chimera of ion pumps and ion channels, with kdpC2 contributing to this specialized mechanism .
Specifically, kdpC2 interacts with the nucleotide-binding loop of KdpB in an ATP-dependent manner, forming a transient KdpB/KdpC/ATP ternary complex. This interaction increases ATP-binding affinity, thereby enhancing the efficiency of the ATP hydrolysis required for potassium transport . This mechanism shares parallels with ABC transporters rather than traditional P-type ATPases, as the ATP nucleotide is coordinated via double hydrogen bonds at a conserved glutamine residue present in KdpC .
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What are the recommended storage and handling conditions for recombinant kdpC2?
For optimal stability and activity of recombinant kdpC2, researchers should follow these storage and handling guidelines:
Storage Conditions:
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Store at -20°C to -80°C for long-term storage
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For working aliquots, store at 4°C for up to one week
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Avoid repeated freeze-thaw cycles, as this can significantly degrade protein quality
Buffer Composition:
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Typically supplied in a Tris-based buffer with 50% glycerol
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pH is optimized for protein stability (usually around pH 8.0)
Handling Recommendations:
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