The loss of inhibitory C-terminal conformations in disease associated P123H β-synuclein.
β-synuclein (βS) is a homologue of α-synuclein (αS), the major protein component of Lewy bodies in patients with Parkinson's disease. In contrast to αS, βS does not form fibrils, mitigates αS toxicity in vivo and inhibits αS fibril formation in vitro. Previously a missense mutation of βS, P123H, was identified in patients with Dementia with Lewy Body disease. The single P123H mutation at the C-terminus of βS is able to convert βS from a nontoxic to a toxic protein that is also able to accelerate formation of inclusions when it is in the presence of αS in vivo. To elucidate the molecular mechanisms of these processes, we compare the conformational properties of the monomer forms of αS, βS and P123H-βS, and the effects on fibril formation of coincubation of αS with βS, and with P123H-βS. NMR residual dipolar couplings and secondary structure propensities show that the P123H mutation of βS renders it more flexible C-terminal to the mutation site and more αS-like. In vitro Thioflavin T fluorescence experiments show that P123H-βS accelerates αS fibril formation upon coincubation, as opposed to wild type βS that acts as an inhibitor of αS aggregation. When P123H-βS becomes more αS-like it is unable to perform the protective function of βS, which suggests that the extended polyproline II motif of βS in the C-terminus is critical to its nontoxic nature and to inhibition of αS upon coincubation. These studies may provide a basis for understanding which regions to target for therapeutic intervention in Parkinson's disease.
Department of Chemistry and Chemical Biology, Rutgers University, Piscataway, New Jersey, 08854.
Maria K
MK
Janowska
Department of Chemistry and Chemical Biology, Rutgers University, Piscataway, New Jersey, 08854.
Jean
J
Baum
D000595:Amino Acid Sequence; D006801:Humans; D007700:Kinetics; D020961:Lewy Body Disease / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D019906:Nuclear Magnetic Resonance, Biomolecular; D010300:Parkinson Disease / Q000235:genetics*; D011487:Protein Conformation; D016415:Sequence Alignment; D051846:beta-Synuclein / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D051846:beta-Synuclein
NMR; P123H-βS; aggregation; dementia with Lewy bodies; fibril formation; inhibition; intrinsically disordered proteins; α-synuclein; β-synuclein