Fast and Easy Transient Mammalian Cell Expression and Purification of Cytoplasmic Dynein.

Abstract

Recombinant protein expression has been key to studying dynein's mechanochemistry and structure-function relationship. To gain further insight into the energy-converting mechanisms and interactions with an increasing variety of dynein cargos and regulators, rapid expression and purification of a variety of dynein proteins and fragments are important. Here we describe transient expression of cytoplasmic dynein in HEK293 cells and fast small-scale purification for high-throughput protein engineering. Mammalian cell expression might be generally considered to be a laborious process, but with recent technology and some simple inexpensive custom-built labware, dynein expression and purification from mammalian cells can be fast and easy.