title: Direct Capture of Functional Proteins from Mammalian Plasma Membranes into Nanodiscs.
abstract: Mammalian plasma membrane proteins make up the largest class of drug targets yet are difficult to study in a cell free system because of their intransigent nature. Herein, we perform direct encapsulation of plasma membrane proteins derived from mammalian cells into a functional nanodisc library. Peptide fingerprinting was used to analyze the proteome of the incorporated proteins in nanodiscs and to further demonstrate that the lipid composition of the nanodiscs directly affects the class of protein that is incorporated. Furthermore, the functionality of the incorporated membrane proteome was evaluated by measuring the activity of membrane proteins: Na(+)/K(+)-ATPase and receptor tyrosine kinases. This work is the first report of the successful establishment and characterization of a cell free functional library of mammalian membrane proteins into nanodiscs.
authors.affiliation: Department of Comparative Biosciences, ‡Department of Veterinary Clinical Medicine, §Department of Chemistry, and ∥Department of Biochemistry, Department of Bioengineering, and Beckman Institute for Advanced Science and Department of Bioengineering, University of Illinois Urbana-Champaign , Urbana, Illinois 61802, United States.
authors.forename: Jahnabi
authors.initials: J
authors.lastname: Roy
authors.affiliation: Department of Comparative Biosciences, ‡Department of Veterinary Clinical Medicine, §Department of Chemistry, and ∥Department of Biochemistry, Department of Bioengineering, and Beckman Institute for Advanced Science and Department of Bioengineering, University of Illinois Urbana-Champaign , Urbana, Illinois 61802, United States.
authors.forename: Holly
authors.initials: H
authors.lastname: Pondenis
authors.affiliation: Department of Comparative Biosciences, ‡Department of Veterinary Clinical Medicine, §Department of Chemistry, and ∥Department of Biochemistry, Department of Bioengineering, and Beckman Institute for Advanced Science and Department of Bioengineering, University of Illinois Urbana-Champaign , Urbana, Illinois 61802, United States.
authors.forename: Timothy M
authors.initials: TM
authors.lastname: Fan
authors.affiliation: Department of Comparative Biosciences, ‡Department of Veterinary Clinical Medicine, §Department of Chemistry, and ∥Department of Biochemistry, Department of Bioengineering, and Beckman Institute for Advanced Science and Department of Bioengineering, University of Illinois Urbana-Champaign , Urbana, Illinois 61802, United States.
authors.forename: Aditi
authors.initials: A
authors.lastname: Das
mesh_terms: D000818:Animals; D002460:Cell Line; D006801:Humans; D008051:Lipid Bilayers / Q000737:chemistry; D008565:Membrane Proteins / Q000302:isolation & purification* / Q000378:metabolism; D008958:Models, Molecular; D049329:Nanostructures / Q000737:chemistry*; D010743:Phospholipids / Q000737:chemistry*; D020543:Proteome / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D008051:Lipid Bilayers; D008565:Membrane Proteins; D010743:Phospholipids; D020543:Proteome