Protein-Engineering - Self assembly - Biochips: Molecular Organization of Proteins in Two Dimensions

Robert Tampé

Celluar Biochemistry and Biophysics, Institute for Physiological Chemistry, Medical School, Karl-von-Frisch-St. 1, D-35043 Marburg, Germany

e-mail: tampe@mailer.uni-marburg.de
 

Due to their self-organizing and dynamic properties, membranes and synthetic amphiphiles produce a wide spectrum of functionality gaining interest in life and material science. These scaffolds offer an ideal tool to organize biomolecules in one or two dimensions that can be manipulated and characterized on nanoscale. In molecular biology, the expression of fusion proteins is a well-established technique of the purification and characterization of gene products. Even a short stretch of surface-exposed histidines enables proteins to specifically interact with metal-chelating complexes. We have combined this approach with the properties of self-organizing systems by the synthesis of metal-chelating lipids and thiols. Proteins can be specifically immobilized and highly oriented at these designed metal-chelating interfaces, eventually leading to two-dimensional protein crystals. These protein arrays are of central interest e.g. for catalytic, switching and sensing biochips and nanodevices.

Literature

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[2] C. Dietrich, L. Schmitt, and R.Tampé, Molecular organization of histidine-tagged biomolecules at self-assembled lipid interfaces using a novel class of chelator lipids. Proc. Natl. Acad. Sci. USA 92, 9014-9018 (1995).

[3] L. Schmitt, T. M. Bohanon, S. Denzinger, H. Ringsdorf, and R. Tampé, Specific protein docking to chelator lipid monolayers monitored by FT-IR at the air-water interface. Angew. Chemie Int. Ed. Engl. 35, 317-320 (1996).

[4] I. T. Dorn, K. Pawlitschko, S. C. Pettinger, and R. Tampé, Orientation and two-dimensional organization of proteins at chelator lipid interfaces. Biol. Chem. 379, 1151-1159. (1998).

[5] I. T. Dorn, K. R. Neumaier, and R. Tampé, Molecular recognition of histidine-tagged molecules by chelator lipids monitored by fluorescence energy transfer and correlation spectroscopy. J. Am. Chem. Soc. 120, 2753-2763. (1998).

[6] I. T. Dorn, R. Escherich, E. Seemüller, R. Guckenberger, and R.Tampé, High-resolution AFM-imaging and mechanistic analysis of the 20 S proteasome. J. Mol. Biol. 288, 1027-1037 (1999).

[7] L. Schmitt, M. Ludwig, H. E. Gaub, and R. Tampé, The molecular force of the NTA/His-tag system - implications for biophysical studies. Biophys. J., in press.