Functional immobilization of a DNA-binding protein at a membrane interface via histidine tag and synthetic chelator lipids. Export

@article{citeulike:3032202, abstract = {The coupling of a DNA-binding protein to self-organized lipid monolayers is examined at the air-water interface by means of film balance techniques and epifluorescence microscopy. We used two recombinant species of the heat shock factor HSF24 which differ only in a carboxy-terminal histidine tag that interacts specifically with the nickel-chelating head group of a synthetic chelator lipid. As key function, HSF24 binds to DNA that contains heat-shock responsible promoter elements. In solution, DNA-protein complex formation is demonstrated for the wild type and fusion protein. Substantial questions of these studies are whether protein function is affected after adsorption to lipid layers and whether a specific docking via histidine tag to the chelator lipid leads to functional immobilization. Using lipid mixtures that allow a lateral organization of chelator lipids within the lipid film, specific binding and unspecific adsorption can be distinguished by pattern formation of DNA-protein complexes. At the lipid interface, functional DNA-protein complexes are only detected, when the histidine-tagged protein was immobilized specifically to a chelator lipid containing monolayer. These results demonstrate that the immobilization of histidine-tagged biomolecules to membranes via chelator lipids is a promising approach to achieve a highly defined deposition of these molecules at an interface maintaining their function.}, address = {Physik Department, Technische Universit\"{a}t M\"{u}nchen, Garching, Germany.}, author = {Dietrich, C. and Boscheinen, O. and Scharf, K. D. and Schmitt, L. and Tamp\'{e}, R.}, citeulike-article-id = {3032202}, citeulike-linkout-0 = {http://dx.doi.org/10.1021/bi952305+}, citeulike-linkout-1 = {http://pubs.acs.org/doi/abs/10.1021/bi952305+}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/8573564}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=8573564}, day = {30}, doi = {10.1021/bi952305+}, issn = {0006-2960}, journal = {Biochemistry}, keywords = {affinity\_tags, complexes, interactions, lipids, methods, monolayer, nickel, purification}, month = {January}, number = {4}, pages = {1100--1105}, posted-at = {2008-07-22 10:07:42}, priority = {2}, title = {Functional immobilization of a DNA-binding protein at a membrane interface via histidine tag and synthetic chelator lipids.}, url = {http://dx.doi.org/10.1021/bi952305+}, volume = {35}, year = {1996} }

TY - JOUR ID - citeulike:3032202 L3 - citeulike-article-id:3032202 N2 - The coupling of a DNA-binding protein to self-organized lipid monolayers is examined at the air-water interface by means of film balance techniques and epifluorescence microscopy. We used two recombinant species of the heat shock factor HSF24 which differ only in a carboxy-terminal histidine tag that interacts specifically with the nickel-chelating head group of a synthetic chelator lipid. As key function, HSF24 binds to DNA that contains heat-shock responsible promoter elements. In solution, DNA-protein complex formation is demonstrated for the wild type and fusion protein. Substantial questions of these studies are whether protein function is affected after adsorption to lipid layers and whether a specific docking via histidine tag to the chelator lipid leads to functional immobilization. Using lipid mixtures that allow a lateral organization of chelator lipids within the lipid film, specific binding and unspecific adsorption can be distinguished by pattern formation of DNA-protein complexes. At the lipid interface, functional DNA-protein complexes are only detected, when the histidine-tagged protein was immobilized specifically to a chelator lipid containing monolayer. These results demonstrate that the immobilization of histidine-tagged biomolecules to membranes via chelator lipids is a promising approach to achieve a highly defined deposition of these molecules at an interface maintaining their function. IS - 4 JF - Biochemistry SN - 0006-2960 AD - Physik Department, Technische Universität München, Garching, Germany. EP - 1105 TI - Functional immobilization of a DNA-binding protein at a membrane interface via histidine tag and synthetic chelator lipids. VL - 35 SP - 1100 KW - affinity_tags KW - complexes KW - interactions KW - lipids KW - methods KW - monolayer KW - nickel KW - purification AU - Dietrich, C AU - Boscheinen, O AU - Scharf, KD AU - Schmitt, L AU - Tampé, R PY - 1996/01/30/ UR - http://dx.doi.org/10.1021/bi952305+ ER -