Production of functional, stable, unmutated recombinant human papillomavirus E6 oncoprotein: Implications for HPV-tumor diagnosis and therapy

摘要: High-risk human papillomaviruses (HR-HPVs) types 16 and 18 are the main etiological agents of cervical cancer, with more than 550,000 new cases each year worldwide. HPVs also associated other ano-genital head-and-neck tumors. The HR-HPV E6 E7 oncoproteins responsible for onset maintenance cell transformation state, they represent appropriate targets development diagnostic therapeutic tools. unmutated gene from HPV16 HPV18 low-risk HPV11 was cloned in a prokaryotic expression vector Histidine-tagged protein (His6-E6), according to novel procedure. structural properties were determined using circular dichroism fluorescence spectroscopy. His6-E6 oncoprotein immunogenicity assessed mouse model, its functionality vitro GST pull-down degradation assays. His6-tagged proteins HPV16, HPV18, genes, without any further modification amino-acid sequence, produced bacteria as soluble stable molecules. Structural analyses suggests that it maintains correct folding conformational properties. C57BL/6 mice immunized developed significant humoral immune responses. purified procedure, investigated protein–protein interactions. bound p53, PDZ1 motif MAGI-1 proteins, discs large tumor suppressor, ubiquitin ligase E6-associated protein, thus suggesting is biologically active. targeted MAGI-3 p53 degradation. This procedure generates stable, which binding will be useful basic studies, kits immunotherapies preclinical models HPV-related tumorigenesis.