title: Targeting the interface of the pathological complex of α-synuclein and TPPP/p25.
abstract: The pathological interaction of intrinsically disordered proteins, such as α-synuclein (SYN) and Tubulin Polymerization Promoting Protein (TPPP/p25), is often associated with neurodegenerative disorders. These hallmark proteins are co-enriched and co-localized in brain inclusions of Parkinson's disease and other synucleinopathies; yet, their successful targeting does not provide adequate effect due to their multiple functions. Here we characterized the interactions of the human recombinant wild type SYN, its truncated forms (SYN(1-120), SYN(95-140)), a synthetized peptide (SYN(126-140)) and a proteolytic fragment (SYN(103-140)) with TPPP/p25 to identify the SYN segment involved in the interaction. The binding of SYN(103-140) to TPPP/p25 detected by ELISA suggested the involvement of a segment within the C-terminal of SYN. The studies performed with ELISA, Microscale Thermophoresis and affinity chromatography proved that SYN(95-140) and SYN(126-140) - in contrast to SYN(1-120) - displayed significant binding to TPPP/p25. Fluorescence assay with ANS, a molten globule indicator, showed that SYN, but not SYN(1-120) abolished the zinc-induced local folding of both the full length as well as the N- and C-terminal-free (core) TPPP/p25; SYN(95-140) and SYN(126-140) were effective as well. The aggregation-prone properties of the SYN species with full length or core TPPP/p25 visualized by immunofluorescent microscopy demonstrated that SYN(95-140) and SYN(126-140), but not SYN(1-120), induced co-enrichment and massive intracellular aggregation after their premixing and uptake from the medium. These data with their innovative impact could contribute to the development of anti-Parkinson drugs with unique specificity by targeting the interface of the pathological TPPP/p25-SYN complex.
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, 1117 Budapest, Hungary. Electronic address: szunyogh.sandor@ttk.mta.hu.
authors.forename: Sándor
authors.initials: S
authors.lastname: Szunyogh
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, 1117 Budapest, Hungary. Electronic address: olah.judit@ttk.mta.hu.
authors.forename: Judit
authors.initials: J
authors.lastname: Oláh
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, 1117 Budapest, Hungary. Electronic address: szenasi.tibor@ttk.mta.hu.
authors.forename: Tibor
authors.initials: T
authors.lastname: Szénási
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, 1117 Budapest, Hungary. Electronic address: szabo.adel@ttk.mta.hu.
authors.forename: Adél
authors.initials: A
authors.lastname: Szabó
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, 1117 Budapest, Hungary. Electronic address: ovadi.judit@ttk.mta.hu.
authors.forename: Judit
authors.initials: J
authors.lastname: Ovádi
keywords: Binding motives; Neomorphic moonlighting function; Parkinson's disease; TPPP/p25; α-Synuclein