TY - JOUR
T1 - Interaction of Alpha-Synuclein and Its Mutants with Rigid Lipid Vesicle Mimics of Varying Surface Curvature
AU - McClain, Sophia M.
AU - Ojoawo, Adedolapo M.
AU - Lin, Wayne
AU - Rienstra, Chad M.
AU - Murphy, Catherine J.
N1 - This work was supported in part by the National Science Foundation (CHE-1608743) and National Institutes of Health (R01-GM112845 and R01-GM123455). S.M.M. is a member of the NIH Chemistry–Biology Interface Training Grant (T32-GM070421). This work was carried out in part in the Materials Research Laboratory Central Facilities, UIUC. We would also like to thank the Protein Sciences Facility of the Roy J. Carver Biotechnology Center, UIUC, for assistance with proteomics analysis, and D. Berthold for assistance in production of protein samples. Portions of this text come from a preliminary version of this work which can be found in the graduate thesis of W.L.
This work was supported in part by the National Science Foundation (CHE-1608743) and National Institutes of Health (R01-GM112845 and R01-GM123455). S.M.M. is a member of the NIH Chemistry?Biology Interface Training Grant (T32-GM070421). This work was carried out in part in the Materials Research Laboratory Central Facilities, UIUC. We would also like to thank the Protein Sciences Facility of the Roy J. Carver Biotechnology Center, UIUC, for assistance with proteomics analysis, and D. Berthold for assistance in production of protein samples. Portions of this text come from a preliminary version of this work which can be found in the graduate thesis of W.L.
PY - 2020/8/25
Y1 - 2020/8/25
N2 - Abnormal aggregation of alpha-synuclein (α-syn), an intrinsically disordered neuronal protein, is strongly implicated in the development of Parkinson's disease. Efforts to better understand α-syn's native function and its pathogenic role in neurodegeneration have revealed that the protein interacts with anionic lipid vesicles via adoption of an amphipathic α-helical structure; however, the ability of α-syn to remodel lipid membranes has made it difficult to decipher the role of vesicle surface curvature in protein binding behavior. In this study, sodium dodecyl sulfate (SDS)-coated gold nanoparticles (AuNPs), which mimic bilayer vesicle architecture, were synthesized in order to conduct a systematic investigation into the binding interaction of α-syn and two of its mutants (A30P and E46K) with rigid lipid vesicle mimics of defined surface curvature. By incorporating a rigid AuNP core (∼10-100 nm), the ability of α-syn to remodel the vesicle mimics was removed and their surface curvature could be fixed. Proteomics studies showed that, upon binding of free α-syn to the surface of SDS-AuNPs, the N-terminus of α-syn became less solvent accessible, whereas its C-terminus became more accessible. Interestingly, α-syn's non-amyloid-β component (NAC) region also exhibited increased solvent accessibility, suggesting that α-syn bound to rigid vesicle-like structures could possess heightened aggregation propensity and therefore pathogenicity. Additionally, both the A30P and E46K mutations were found to adopt distinct binding modes on the mimics' surface. In contrast with previous reports, similar binding affinities were observed for WT, A30P, and E46K α-syn toward SDS-AuNPs of all sizes, indicating the potential importance of vesicle deformability in determining α-syn binding behavior.
AB - Abnormal aggregation of alpha-synuclein (α-syn), an intrinsically disordered neuronal protein, is strongly implicated in the development of Parkinson's disease. Efforts to better understand α-syn's native function and its pathogenic role in neurodegeneration have revealed that the protein interacts with anionic lipid vesicles via adoption of an amphipathic α-helical structure; however, the ability of α-syn to remodel lipid membranes has made it difficult to decipher the role of vesicle surface curvature in protein binding behavior. In this study, sodium dodecyl sulfate (SDS)-coated gold nanoparticles (AuNPs), which mimic bilayer vesicle architecture, were synthesized in order to conduct a systematic investigation into the binding interaction of α-syn and two of its mutants (A30P and E46K) with rigid lipid vesicle mimics of defined surface curvature. By incorporating a rigid AuNP core (∼10-100 nm), the ability of α-syn to remodel the vesicle mimics was removed and their surface curvature could be fixed. Proteomics studies showed that, upon binding of free α-syn to the surface of SDS-AuNPs, the N-terminus of α-syn became less solvent accessible, whereas its C-terminus became more accessible. Interestingly, α-syn's non-amyloid-β component (NAC) region also exhibited increased solvent accessibility, suggesting that α-syn bound to rigid vesicle-like structures could possess heightened aggregation propensity and therefore pathogenicity. Additionally, both the A30P and E46K mutations were found to adopt distinct binding modes on the mimics' surface. In contrast with previous reports, similar binding affinities were observed for WT, A30P, and E46K α-syn toward SDS-AuNPs of all sizes, indicating the potential importance of vesicle deformability in determining α-syn binding behavior.
KW - SDS
KW - alpha-synuclein
KW - gold nanoparticles
KW - limited proteolysis
KW - lipid vesicle mimics
KW - surface curvature
UR - https://www.scopus.com/pages/publications/85090078386
UR - https://www.scopus.com/pages/publications/85090078386#tab=citedBy
U2 - 10.1021/acsnano.0c03420
DO - 10.1021/acsnano.0c03420
M3 - Article
C2 - 32672441
AN - SCOPUS:85090078386
SN - 1936-0851
VL - 14
SP - 10153
EP - 10167
JO - ACS Nano
JF - ACS Nano
IS - 8
ER -