Display cell surface

Figure 3.3 Cell surface display. Proteins displayed on the cell surface of organisms bind a fluorescent molecule. Cells are passed through a FACS machine, allowing separation into populations that do or do not bind the target...

Becker, S., Schmoldt, H.U., Adams, T.M. et al. (2004) Ultra-high-throughput screening based on cell-surface display and fluorescence-activated cell sorting for the identification of novel biocatalysts. Current Opinion in Biotechnology, 15, 323-329. 

Shimazu, M., Mulchandani, A. and Chen, W. (2001) Cell surface display of organophosphorus hydrolase using ice nucleation protein. Biotechnology Progress, 17, 76-80. 

Daugherty, P.S. and Morse, D.E. (2005) Enzymatic synthesis of layered titanium phosphates at low temperature and neutral pH by cell-surface display of silicatein a. Journal of the American Chemical Society, 127, 15749-15755. 

From the information given above it is obvious that cell surfaces display an enormous complexity. A perfect model to study the interaction of a peptide with a biological membrane would require knowledge about the cell membrane composition in that particular tissue. Even if such information were available it will most probably not be possible to fully mimic the biological environment. However, some important aspects may still be studied with the available models. Whenever possible, one should try to relate the information derived from such a model to information gained from biological data taken on real cells (cell-lines) such as binding affinities etc. in order to prove the validity of the model for the study of a particular aspect. 

Yarema, K.J. 2001. New directions in carbohydrate engineering a metabolic substrate-based approach to modify the cell surface display of sialic acids. Bio techniques 31 384-393. 

Successful cell surface display of the protein can be verified by inducing gene expression via addition of anhydrotetracycline to the culture medium and immunofluorescence staining of the cells using an antibody directed against the protein to be displayed (Protocol 1). Analysis can be performed by fluorescence microscopy or flow cytometry. 

Ligand binding, and certain enzymatic assays, can be greatly simplified if a protein is biosynthetically anchored on the external surface of a cell. Surface displayed proteins are readily accessible to fluorescent probes and thus, transport limitations are circumvented. In addition,... 

Christmann, A., Walter, K., Wentzel, A., Kratzner, R., and Kolmar, H. (1999). The cystine knot of a squash-type protease inhibitor as a structural scaffold for Escherichia coli cell surface display of conformationally constrained peptides. Prot. Eng. 12, 797-806. 

Nguyen TN, Hansson M, Stahl S, Bachi T, Robert A, Domzig W, Binz H, Uhlen M, Cell-surface display of heterologous epitopes on Staphylococcus xvlosus as a potential delivery system for oral vaccination, Gene, 128 89-94, 1993. 

Directed Evolution of Binding Proteins by Cell Surface Display Analysis of the Screening Process... 

O Section 3 of this chapter described strategies based on the metabolic engineering of several monosaccharides to achieve cell surface display of chemical functional groups including... 

Ketone-based chemoselective ligation reactions that exploit cell surface displayed SiaSLev (32)... 

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