Functional Surface Attachment in a Sandwich Geometry of GFP-Labeled Motor Proteins
Molecular motors perform work in cells by moving in an ATP-dependent manner along filamentous tracks. In vitro, the mechanical action of such motor proteins can be investigated by attaching the molecules to surfaces in the so-called gliding or bead assays. Surface attachment protocols have to be used that do not interfere with the function of the molecule. Here, we describe a sandwich protocol that preserves functionality. The protocol can be used for a large variety of proteins, in particular kinesin motor proteins that are GFP-tagged.
Key wordsMotor protein Surface attachment Kinesin Kip3p Gliding assay Bead assay
The authors acknowledge the input of members of the Howard, Schäffer, and Diez labs in the development of these protocols.
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