title: Solid-state NMR spectroscopy of protein complexes.
abstract: Protein-protein interactions are vital for many biological processes. These interactions often result in the formation of protein assemblies that are large in size, insoluble, and difficult to crystallize, and therefore are challenging to study by structure biology techniques, such as single crystal X-ray diffraction and solution NMR spectroscopy. Solid-state NMR (SSNMR) spectroscopy is emerging as a promising technique for studies of such protein assemblies because it is not limited by molecular size, solubility, or lack of long-range order. In the past several years, we have applied magic angle spinning SSNMR-based methods to study several protein complexes. In this chapter, we discuss the general SSNMR methodologies employed for structural and dynamics analyses of protein complexes with specific examples from our work on thioredoxin reassemblies, HIV-1 capsid protein assemblies, and microtubule-associated protein assemblies. We present protocols for sample preparation and characterization, pulse sequences, SSNMR spectra collection, and data analysis.
authors.affiliation: Department of Chemistry and Biochemistry, University of Delaware, Newark, DE, USA.
authors.forename: Shangjin
authors.initials: S
authors.lastname: Sun
authors.forename: Yun
authors.initials: Y
authors.lastname: Han
authors.forename: Sivakumar
authors.initials: S
authors.lastname: Paramasivam
authors.forename: Si
authors.initials: S
authors.lastname: Yan
authors.forename: Amanda E
authors.initials: AE
authors.lastname: Siglin
authors.forename: John C
authors.initials: JC
authors.lastname: Williams
authors.forename: In-Ja L
authors.initials: IJ
authors.lastname: Byeon
authors.forename: Jinwoo
authors.initials: J
authors.lastname: Ahn
authors.forename: Angela M
authors.initials: AM
authors.lastname: Gronenborn
authors.forename: Tatyana
authors.initials: T
authors.lastname: Polenova
mesh_terms: D036022:Capsid Proteins / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D002247:Carbon Isotopes / Q000378:metabolism; D020285:Cryoelectron Microscopy / Q000379:methods; D015497:HIV-1 / Q000737:chemistry / Q000648:ultrastructure; D018613:Microscopy, Confocal / Q000379:methods; D046529:Microscopy, Electron, Transmission / Q000379:methods; D008868:Microtubule Proteins / Q000737:chemistry / Q000378:metabolism; D046912:Multiprotein Complexes / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D009587:Nitrogen Isotopes / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D013879:Thioredoxins / Q000737:chemistry / Q000378:metabolism
chemical_list: D036022:Capsid Proteins; D002247:Carbon Isotopes; D008868:Microtubule Proteins; D046912:Multiprotein Complexes; D009587:Nitrogen Isotopes; D013879:Thioredoxins