FN3K Human
Description
FN3K is fused to a 23 amino acid His-tag at N-terminus & purified by proprietary chromatographic techniques.
Product Specs
Q&A
Experimental Design for Studying FN3K Inhibitors
Q: How can researchers design experiments to screen and evaluate the efficacy of FN3K inhibitors in human cells? A: To screen FN3K inhibitors, researchers can employ a multi-step approach:
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Homology Modeling: Use tools like SWISS Expasy to create a 3D model of human FN3K, as its structure is not available in databases like PDB .
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Virtual Screening: Utilize molecular docking to identify potential inhibitors that interact with the catalytic domain of FN3K .
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Molecular Dynamics Simulations: Validate the stability and interactions of inhibitors with FN3K .
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Cytotoxicity Assays: Test the efficacy and safety of inhibitors in cell lines like BT-474 and T-47D .
Understanding FN3K's Role in Glycation and Disease
Q: What is the role of FN3K in protein deglycation, and how does it relate to diseases like diabetes and cancer? A: FN3K phosphorylates fructosamines, facilitating the removal of glycated proteins. This process is crucial in conditions like diabetes and cancer, where glycation impacts protein function and cellular stress . FN3K's activity helps maintain cellular homeostasis by repairing glycated proteins, which can otherwise contribute to oxidative stress and disease progression .
Analyzing Data Contradictions in FN3K Research
Q: How can researchers address discrepancies in data regarding FN3K's substrate specificity and its implications for therapeutic applications? A: Discrepancies in substrate specificity can arise from differences in experimental conditions or assay sensitivity. Researchers should:
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Validate Assays: Ensure that assays are specific and sensitive to detect FN3K activity accurately.
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Compare Conditions: Standardize experimental conditions across studies to ensure comparability.
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Consider Orthologs: Study FN3K orthologs, like FN3KRP, which may have different substrate specificities, to understand the evolutionary conservation of FN3K's function .
Advanced Research Questions on FN3K Structure and Function
Q: What insights do recent structural studies of human FN3K provide for understanding its catalytic mechanism and potential therapeutic targeting? A: Recent crystal structures of human FN3K reveal key features for its kinase activity and substrate recognition . These structures provide a basis for designing small molecule inhibitors targeting the FN3K-NRF2 axis, which is crucial in cancer and oxidative stress regulation .
Methodological Considerations for FN3K Inhibition Studies
Q: What methodological considerations are essential when designing studies to inhibit FN3K activity in cellular models? A: When studying FN3K inhibition:
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Use Appropriate Cell Models: Choose cell lines relevant to the disease context, such as breast cancer cells for cancer studies .
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Validate Inhibitor Specificity: Ensure that inhibitors are specific to FN3K and do not affect other kinases.
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Assess Cellular Effects: Monitor changes in cellular stress markers and protein deglycation levels upon FN3K inhibition .
Integrating FN3K into Broader Biological Pathways
Q: How does FN3K interact with other cellular pathways, particularly those involved in oxidative stress and cancer? A: FN3K interacts with pathways involving NRF2, a key regulator of oxidative stress. By deglycating NRF2, FN3K modulates its activity, influencing antioxidant signaling and potentially impacting cancer cell survival . Understanding these interactions is crucial for developing therapeutic strategies targeting FN3K in cancer.
Future Directions in FN3K Research
Q: What are some future research directions for FN3K, especially in the context of disease prevention and treatment? A: Future research should focus on:
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Therapeutic Inhibitor Design: Developing specific inhibitors for FN3K to modulate its activity in disease contexts .
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Mechanistic Studies: Elucidating the detailed mechanisms of FN3K in protein deglycation and its impact on cellular stress pathways .
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Translational Studies: Investigating the potential of FN3K as a therapeutic target in clinical settings .
Data Analysis and Validation in FN3K Studies
Q: How can researchers validate and analyze data from FN3K studies to ensure reliability and reproducibility? A: To validate data:
Comparative Analysis of FN3K Across Species
Q: What insights can be gained from comparing FN3K function and structure across different species? A: Comparing FN3K across species, such as between humans and Arabidopsis thaliana, can provide insights into evolutionary conservation and functional divergence. This comparison can help identify conserved motifs essential for FN3K activity and substrate specificity .
Implications of FN3K Research for Clinical Applications
Q: How might research on FN3K impact clinical practice, particularly in managing conditions like diabetes and cancer? A: Understanding FN3K's role in protein deglycation could lead to novel therapeutic strategies for diseases where glycation is a factor. By modulating FN3K activity, it may be possible to mitigate oxidative stress and improve outcomes in diabetes and cancer . Clinical trials focusing on FN3K inhibitors could provide valuable insights into their efficacy and safety in human patients.
Table: Key Features of Human FN3K and Its Implications
Figure: FN3K's Role in Protein Deglycation
FN3K phosphorylates the fructose-lysine moiety, leading to the removal of glycated proteins. This process involves the conversion of the Schiff base intermediate into a harmless ketoaldehyde, restoring the protein to its non-glycated state .
Product Science Overview
The primary function of FN3K is to phosphorylate fructoselysine residues on glycated proteins, converting them to fructoselysine-3-phosphate (FL3P). This phosphorylation destabilizes the FL adduct, leading to its spontaneous decomposition and effectively reversing the non-enzymatic glycation process at an early stage . This process is vital for protecting cells from the deleterious effects of non-enzymatic glycation, which is a significant factor in the pathogenesis of diabetic complications .
The concentrations of fructosamines and advanced glycation end products (AGEs) are elevated in diabetes, contributing to various complications . FN3K’s ability to deglycate these compounds makes it an important enzyme in mitigating the effects of diabetes. Loss of FN3K’s protective enzymatic activity has also been associated with the oncology of colorectal cancer .
Recombinant human FN3K is produced using E. coli expression systems. The recombinant protein is typically supplied as a carrier-free solution, which enhances protein stability and increases shelf-life . It is used in various research applications, including studies on diabetes and protein glycation.
