Self-assembled monolayers improve protein distribution on holey carbon cryo-EM supports.
Poor partitioning of macromolecules into the holes of holey carbon support grids frequently limits structural determination by single particle cryo-electron microscopy (cryo-EM). Here, we present a method to deposit, on gold-coated carbon grids, a self-assembled monolayer whose surface properties can be controlled by chemical modification. We demonstrate the utility of this approach to drive partitioning of ionotropic glutamate receptors into the holes, thereby enabling 3D structural analysis using cryo-EM methods.
Laboratory of Cell Biology, Center for Cancer Research, NCI, NIH, Bethesda, MD 20892.
Joel R
JR
Meyerson
Laboratory of Cell Biology, Center for Cancer Research, NCI, NIH, Bethesda, MD 20892.
Prashant
P
Rao
Laboratory of Cellular and Molecular Neurophysiology, Porter Neuroscience Research Center, NICHD, NIH, Bethesda MD 20892.
Janesh
J
Kumar
Laboratory of Cellular and Molecular Neurophysiology, Porter Neuroscience Research Center, NICHD, NIH, Bethesda MD 20892.
Sagar
S
Chittori
Laboratory of Cell Biology, Center for Cancer Research, NCI, NIH, Bethesda, MD 20892.
Soojay
S
Banerjee
FEI Company, Hillsboro, OR 97124.
Jason
J
Pierson
Laboratory of Cellular and Molecular Neurophysiology, Porter Neuroscience Research Center, NICHD, NIH, Bethesda MD 20892.
Mark L
ML
Mayer
Laboratory of Cell Biology, Center for Cancer Research, NCI, NIH, Bethesda, MD 20892.
Sriram
S
Subramaniam
D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry*; D002244:Carbon / Q000737:chemistry; D018834:Chaperonin 60 / Q000737:chemistry*; D020285:Cryoelectron Microscopy / Q000295:instrumentation*; D006046:Gold / Q000737:chemistry; D016062:Porosity; D051381:Rats; D018091:Receptors, AMPA / Q000737:chemistry*; D018092:Kainic Acid Receptors / Q000737:chemistry*; D017155:Wettability; D000097806:GluK2 Kainate Receptor
D001426:Bacterial Proteins; D002244:Carbon; D018834:Chaperonin 60; D006046:Gold; D018091:Receptors, AMPA; D018092:Kainic Acid Receptors; D000097806:GluK2 Kainate Receptor; C104722:glutamate receptor ionotropic, AMPA 2