tailieunhanh - báo cáo khoa học: "Binary polypeptide system for permanent and oriented protein immobilization"

Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: Binary polypeptide system for permanent and oriented protein immobilization | Ferrari et al. Journal of Nanobiotechnology 2010 8 9 http content 8 1 9 RESEARCH JOURNAL OF NANOBIOTECHNOLOGY Open Access Binary polypeptide system for permanent and oriented protein immobilization Enrico Ferrari 1 Frédéric Darios1 Fan Zhang1 Dhevahi Niranjan1 Julian Bailes2 Mikhail Soloviev2 and Bazbek Davletov 1 Abstract Background Many techniques in molecular biology clinical diagnostics and biotechnology rely on binary affinity tags. The existing tags are based on either small molecules . biotin streptavidin or glutathione GST or peptide tags FLAG Myc HA Strep-tag and His-tag . Among these the biotin-streptavidin system is most popular due to the nearly irreversible interaction of biotin with the tetrameric protein streptavidin. The major drawback of the stable biotinstreptavidin system however is that neither of the two tags can be added to a protein of interest via recombinant means except for the Strep-tag case leading to the requirement for chemical coupling. Results Here we report a new immobilization system which utilizes two monomeric polypeptides which selfassemble to produce non-covalent yet nearly irreversible complex which is stable in strong detergents chaotropic agents as well as in acids and alkali. Our system is based on the core region of the tetra-helical bundle known as the SNARE soluble N-ethylmaleimide-sensitive factor attachment protein receptor complex. This irreversible protein attachment system IPAS uses either a shortened syntaxin helix and fused SNAP25-synaptobrevin or a fused syntaxin-synaptobrevin and SNAP25 allowing a two-component system suitable for recombinant protein tagging capture and immobilization. We also show that IPAS is suitable for use with traditional beads and chromatography planar surfaces and Biacore gold nanoparticles and for protein-protein interaction in solution. Conclusions IPAS offers an alternative to chemical cross-linking streptavidin-biotin system and to traditional peptide .