Reconstitution of full-length human caveolin-1 into phospholipid bicelles: Validation by analytical ultracentrifugation.

作者: Monica D. Rieth , Kyle T. Root , Kerney Jebrell Glover

DOI: 10.1016/J.BPC.2020.106339

关键词:

摘要: A significant hurdle in obtaining biophysical information on membrane proteins is developing a successful strategy for their reconstitution into suitable mimic. In particular, utilization of the more 'native-like' mimics such as bicelles generally challenging than simple micellar solubilization. Caveolin-1, an integral protein involved curvature, endocytosis, mechano-protection, and signal transduction, has been shown to be particularly recalcitrant standard protocols due its highly hydrophobic characteristics. Herein we describe robust method incorporate recombinantly produced full-length caveolin-1 at levels needed experimentation. The benchmark obtainment homogeneous state; therefore, developed validation procedure monitor success using analytical ultracentrifugation density-matched bicelles. Our findings indicated that our protocol produces very preparation associated with bicelles, α-helical (by circular dichroism spectroscopy). We believe this methodology will serve general facilitate studies proteins.

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