NMR Structure of Carcinoscorpius rotundicauda Thioredoxin-related Protein 16 and Its Role in Regulating Transcription Factor NF-κB Activity

Abstract

Background: Cr-TRP16 is a thioredoxin like protein that regulates NF-B activity. Results: NMR structure of Cr-TRP16, along with its regulation of NF-B activity, is reported. Conclusion: Our findings elucidate molecular mechanism by which NF-B activation is regulated through Cr-TRP16. Significance: This study helps further in understanding the interactions of Trxs and Trx-like proteins with other metabolic pathways and will lead to promising therapeutic development.

Cooperatively transcriptional and epigenetic regulation of sonic hedgehog overexpression drives malignant potential of breast cancer

“…To determine NF‐κB activation, we performed a DNA binding assay using the TransAM NF‐κB kit (Active Motif, Carlsbad, CA, USA) according to the manufacturer's instructions and as previously described . Briefly, MDA‐MB‐231 cells were treated with 5‐Aza (20 μM) or TNF‐α (20 ng/mL) for 24 h. Nuclear protein extracts were prepared using a nuclear extraction kit (Invitrogen) according to the manufacturer's instructions.…”

“…To determine NF-jB activation, we performed a DNA binding assay using the TransAM NF-jB kit (Active Motif, Carlsbad, CA, USA) according to the manufacturer's instructions and as previously described. (24) Briefly, MDA-MB-231 cells were treated with 5-Aza (20 lM) or TNF-a (20 ng ⁄ mL) for 24 h. Nuclear protein extracts were prepared using a nuclear extraction kit (Invitrogen) according to the manufacturer's instructions. Protein concentration was determined using the BCA Protein Assay Kit (Pierce), and 20 lg of nuclear extract was added to the plates precoated with an NF-jB consensus oligonucleotide sequence and incubated at room temperature for 1 h with continuous shaking.…”

Thioredoxin-like protein 2b facilitates colon cancer cell proliferation and inhibits apoptosis via NF-κB pathway

“…Interestingly, NF-κB is a redox-sensitive factor. Many Trx members reduce the Cys residue in the DNA-binding motif of NF-κB to enhance its binding to target promoter sites, which is more active than nonphysiological reducing agents [30,31].…”