Article ID Journal Published Year Pages File Type
2828998 Journal of Structural Biology 2009 6 Pages PDF
Abstract

Crystalline bacterial cell surface layers (S-layers) show the ability to recrystallize into highly regular pattern on solid supports. In this study, the genetically modified S-layer protein SbpA of Lysinibacillus sphaericus CCM 2177, carrying a hexa-histidine tag (His6-tag) at the C-terminus, was used to generate functionalized two-dimensional nanoarrays on a silicon surface. Atomic force microscopy (AFM) was applied to explore the topography and the functionality of the fused His6-tags. The accessibility of the His6-tags was demonstrated by in-situ anti-His-tag antibody binding to the functional S-layer array. The metal binding properties of the His6-tag was investigated by single molecule force microscopy. For this purpose, newly developed tris–NTA was tethered to the AFM tips via a flexible polyethylene glycol (PEG) linker. The functionalized tips showed specific interactions with S-layer containing His6-tags in the presence of nickel ions. Thus the His6-tag is located at the outer surface of the S-layer and can be used for stable but reversible attachment of functional tris–NTA derivatives.

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Life Sciences Biochemistry, Genetics and Molecular Biology Molecular Biology
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