title: Facile backbone structure determination of human membrane proteins by NMR spectroscopy.
abstract: Although nearly half of today's major pharmaceutical drugs target human integral membrane proteins (hIMPs), only 30 hIMP structures are currently available in the Protein Data Bank, largely owing to inefficiencies in protein production. Here we describe a strategy for the rapid structure determination of hIMPs, using solution NMR spectroscopy with systematically labeled proteins produced via cell-free expression. We report new backbone structures of six hIMPs, solved in only 18 months from 15 initial targets. Application of our protocols to an additional 135 hIMPs with molecular weight <30 kDa yielded 38 hIMPs suitable for structural characterization by solution NMR spectroscopy without additional optimization.
authors.affiliation: Structural Biology Laboratory, Salk Institute for Biological Studies, La Jolla, California, USA.
authors.forename: Christian
authors.initials: C
authors.lastname: Klammt
authors.forename: Innokentiy
authors.initials: I
authors.lastname: Maslennikov
authors.forename: Monika
authors.initials: M
authors.lastname: Bayrhuber
authors.forename: Cédric
authors.initials: C
authors.lastname: Eichmann
authors.forename: Navratna
authors.initials: N
authors.lastname: Vajpai
authors.forename: Ellis Jeremy Chua
authors.initials: EJ
authors.lastname: Chiu
authors.forename: Katherine Y
authors.initials: KY
authors.lastname: Blain
authors.forename: Luis
authors.initials: L
authors.lastname: Esquivies
authors.forename: June Hyun Jung
authors.initials: JH
authors.lastname: Kwon
authors.forename: Bartosz
authors.initials: B
authors.lastname: Balana
authors.forename: Ursula
authors.initials: U
authors.lastname: Pieper
authors.forename: Andrej
authors.initials: A
authors.lastname: Sali
authors.forename: Paul A
authors.initials: PA
authors.lastname: Slesinger
authors.forename: Witek
authors.initials: W
authors.lastname: Kwiatkowski
authors.forename: Roland
authors.initials: R
authors.lastname: Riek
authors.forename: Senyon
authors.initials: S
authors.lastname: Choe
mesh_terms: D030562:Databases, Protein; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry*; D008958:Models, Molecular; D008970:Molecular Weight; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011487:Protein Conformation
chemical_list: D008565:Membrane Proteins