ESCORTing proteins directly from whole cell-lysate for single-molecule studies

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Abstract

We have developed a method for Enzymatic Sortase-assisted Covalent Orientation-specific Restraint Tethering (ESCORT) recombinant proteins onto surfaces directly from cell-lysate. With an improved surface passivation method, we obviate the cumbersome purification steps even for single molecule studies that demand high purity in the sample. We demonstrated high-specificity of the method, high-passivity of the surface and uncompromised functional integrity of anchored proteins using single molecule fluorescence and force-mapping. We anticipate that this method will substantially reduce the investment by way of time, money and energy in the area of single molecule studies.

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Srinivasan, S., Hazra, J. P., Singaraju, G. S., Deb, D., & Rakshit, S. (2017). ESCORTing proteins directly from whole cell-lysate for single-molecule studies. Analytical Biochemistry, 535, 35–42. https://doi.org/10.1016/j.ab.2017.07.022

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