TY - JOUR
T1 - DNP-Enhanced MAS NMR
T2 - A Tool to Snapshot Conformational Ensembles of α-Synuclein in Different States
AU - Uluca, Boran
AU - Viennet, Thibault
AU - Petrović, Dušan
AU - Shaykhalishahi, Hamed
AU - Weirich, Franziska
AU - Gönülalan, Ayşenur
AU - Strodel, Birgit
AU - Etzkorn, Manuel
AU - Hoyer, Wolfgang
AU - Heise, Henrike
N1 - Publisher Copyright:
© 2018 Biophysical Society
PY - 2018/4/10
Y1 - 2018/4/10
N2 - Intrinsically disordered proteins dynamically sample a wide conformational space and therefore do not adopt a stable and defined three-dimensional conformation. The structural heterogeneity is related to their proper functioning in physiological processes. Knowledge of the conformational ensemble is crucial for a complete comprehension of this kind of proteins. We here present an approach that utilizes dynamic nuclear polarization-enhanced solid-state NMR spectroscopy of sparsely isotope-labeled proteins in frozen solution to take snapshots of the complete structural ensembles by exploiting the inhomogeneously broadened line-shapes. We investigated the intrinsically disordered protein α-synuclein (α-syn), which plays a key role in the etiology of Parkinson's disease, in three different physiologically relevant states. For the free monomer in frozen solution we could see that the so-called “random coil conformation” consists of α-helical and β-sheet-like conformations, and that secondary chemical shifts of neighboring amino acids tend to be correlated, indicative of frequent formation of secondary structure elements. Based on these results, we could estimate the number of disordered regions in fibrillar α-syn as well as in α-syn bound to membranes in different protein-to-lipid ratios. Our approach thus provides quantitative information on the propensity to sample transient secondary structures in different functional states. Molecular dynamics simulations rationalize the results.
AB - Intrinsically disordered proteins dynamically sample a wide conformational space and therefore do not adopt a stable and defined three-dimensional conformation. The structural heterogeneity is related to their proper functioning in physiological processes. Knowledge of the conformational ensemble is crucial for a complete comprehension of this kind of proteins. We here present an approach that utilizes dynamic nuclear polarization-enhanced solid-state NMR spectroscopy of sparsely isotope-labeled proteins in frozen solution to take snapshots of the complete structural ensembles by exploiting the inhomogeneously broadened line-shapes. We investigated the intrinsically disordered protein α-synuclein (α-syn), which plays a key role in the etiology of Parkinson's disease, in three different physiologically relevant states. For the free monomer in frozen solution we could see that the so-called “random coil conformation” consists of α-helical and β-sheet-like conformations, and that secondary chemical shifts of neighboring amino acids tend to be correlated, indicative of frequent formation of secondary structure elements. Based on these results, we could estimate the number of disordered regions in fibrillar α-syn as well as in α-syn bound to membranes in different protein-to-lipid ratios. Our approach thus provides quantitative information on the propensity to sample transient secondary structures in different functional states. Molecular dynamics simulations rationalize the results.
UR - http://www.scopus.com/inward/record.url?scp=85044971717&partnerID=8YFLogxK
U2 - 10.1016/j.bpj.2018.02.011
DO - 10.1016/j.bpj.2018.02.011
M3 - Journal article
C2 - 29642031
AN - SCOPUS:85044971717
SN - 0006-3495
VL - 114
SP - 1614
EP - 1623
JO - Biophysical Journal
JF - Biophysical Journal
IS - 7
ER -