title: Differential dynamical effects of macromolecular crowding on an intrinsically disordered protein and a globular protein: implications for in-cell NMR spectroscopy.
abstract: In-cell NMR provides a valuable means to assess how macromolecules, with concentrations up to 300 g/L in the cytoplasm, affect the structure and dynamics of proteins at atomic resolution. Here an intrinsically disordered protein, alpha-synuclein (alphaSN), and a globular protein, chymotrypsin inhibitor 2 (CI2) were examined by using in-cell NMR. High-resolution in-cell spectra of alphaSN can be obtained, but CI2 leaks from the cell and the remaining intracellular CI2 is not detectable. Even after stabilizing the cells from leakage by using alginate encapsulation, no CI2 signal is detected. From in vitro studies we conclude that this difference in detectability is the result of the differential dynamical response of disordered and ordered proteins to the changes of motion caused by the increased viscosity in cells.
authors.affiliation: Department of Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.
authors.forename: Conggang
authors.initials: C
authors.lastname: Li
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Charlton
authors.forename: Asha
authors.initials: A
authors.lastname: Lakkavaram
authors.forename: Christopher
authors.initials: C
authors.lastname: Seagle
authors.forename: Guifang
authors.initials: G
authors.lastname: Wang
authors.forename: Gregory B
authors.initials: GB
authors.lastname: Young
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Macdonald
authors.forename: Gary J
authors.initials: GJ
authors.lastname: Pielak
mesh_terms: D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010455:Peptides / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry*; D011205:Povidone / Q000737:chemistry*; D014783:Viscosity; D051844:alpha-Synuclein / Q000737:chemistry*
chemical_list: D010455:Peptides; D010940:Plant Proteins; D051844:alpha-Synuclein; C093336:chymotrypsin inhibitor 2; D011205:Povidone