Identification of motives mediating alternative functions of the neomorphic moonlighting TPPP/p25

Tõkési, Natália; Oláh, Judit; Hlavanda, Emma; Szunyogh, Sándor; Szabó, Adél; Babos, Fruzsina; Magyar, Anna; Lehotzky, Attila; Vass, Elemér; Ovádi, Judit

doi:10.1016/j.bbadis.2014.01.009

Cited by 25 publications

(78 citation statements)

References 35 publications

(62 reference statements)

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“…Earlier we showed that the 147 KAPIISGVTK 156 segment of the flexible CORE region of TPPP/p25 is involved in the formation of the pathological TPPP/p25-SYN complex, while the 178-187 segment of the C-terminus takes part in the formation of the physiological TPPP/ p25-tubulin/microtubule complex [35]. Deletion and/or truncated variants and fragments of the human TPPP/p25 were produced by recombinant techniques as described in the Materials and Methods and used for structural and functional studies.…”

Section: Generation Of Deletion Mutants Of Tppp/p25mentioning

confidence: 99%

“…Recently we have suggested a new strategy for the therapeutic treatment of Parkinson's disease by targeting the interface of the pathological TPPP/p25-SYN complex without influencing the physiological interaction of TPPP/p25 with tubulin/microtubules [35,36]. The distinct interface of the physiological and pathological complexes is a crucial point in our drug targeting strategy.…”

Section: U N C O R R E C T E D P R O O Fmentioning

confidence: 99%

“…Δ3-43/Δ175-219, DT) was prepared and purified as described previously [35]. The double truncated mini loop TPPP/p25 (DTML), double truncated small loop TPPP/p25 (DTSL) and double truncated double loop TPPP/p25 (DTDL) were amplified by PCR using the following primer pairs: primer13 and primer17 and ML, SL or DL as a template.…”

Section: Dna Manipulationsmentioning

confidence: 99%

“…Human recombinant TPPP/p25 forms and fragments (see Table 2) possessing His-tags were expressed in E. coli BL21 (DE3) cells and isolated on HIS-Select™ Cartridge (Sigma-Aldrich) as described previously [16,35].…”

Section: Expression and Purification Of Human Recombinant Wild Type Amentioning

confidence: 99%

“…The plate was coated with 5 μg/ml (50 μl/well) SYN in phosphate buffered saline as described previously [35]. Briefly, after blocking the wells with bovine serum albumin (BSA), the immobilized proteins were incubated with serial dilutions of wild type or mutant TPPP/p25 forms followed by the addition of TPPP/p25 antibody [16] and the corresponding peroxidase conjugated secondary IgG.…”

Section: Enzyme-linked Immunosorbent Assay (Elisa)mentioning

confidence: 99%

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Challenging drug target for Parkinson's disease: Pathological complex of the chameleon TPPP/p25 and alpha-synuclein proteins

Szénási¹,

Oláh²,

Szabó³

et al. 2017

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Diseas

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The hallmarks of Parkinson's disease and other synucleinopathies, Tubulin Polymerization Promoting Protein (TPPP/p25) and α-synuclein (SYN) have two key features: they are disordered and co-enriched/co-localized in brain inclusions. These Neomorphic Moonlighting Proteins display both physiological and pathological functions due to their interactions with distinct partners. To achieve the selective targeting of the pathological TPPP/p25-SYN but not the physiological TPPP/ p25-tubulin complex, their interfaces were identified as a specific innovative strategy for the development of anti-Parkinson drugs. Therefore, the interactions of TPPP/p25 with tubulin and SYN were characterized which suggested the involvements of the 178-187 aa and 147-156 aa segments in the complexation of TPPP/p25 with tubulin and SYN, respectively. However, various truncated and deletion mutants reduced but did not abolish the interactions except one mutant; in addition synthetized fragments corresponding to the potential binding segments of TPPP/p25 failed to interact with SYN. In fact, the studies of the multiple interactions at molecular and cellular levels revealed the high conformational plasticity, chameleon feature, of TPPP/p25 that ensures exceptional functional resilience; the lack of previously identified binding segments could be replaced by other segments. The experimental results are underlined by distinct bioinformatics tools. All these data revealed that although targeting chameleon proteins is a challenging task, nevertheless, the validation of a drug target can be achieved by identifying the interface of complexes of the partner proteins existing at the given pathological conditions.

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Section: Generation Of Deletion Mutants Of Tppp/p25mentioning

confidence: 99%

Section: U N C O R R E C T E D P R O O Fmentioning

confidence: 99%

Section: Dna Manipulationsmentioning

confidence: 99%

Section: Expression and Purification Of Human Recombinant Wild Type Amentioning

confidence: 99%

Section: Enzyme-linked Immunosorbent Assay (Elisa)mentioning

confidence: 99%

See 3 more Smart Citations

Challenging drug target for Parkinson's disease: Pathological complex of the chameleon TPPP/p25 and alpha-synuclein proteins

Szénási¹,

Oláh²,

Szabó³

et al. 2017

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Diseas

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Pharmacological targeting of α‐synuclein and TPPP/p25 in Parkinson's disease: challenges and opportunities in a Nutshell

Oláh

Ovádi

2019

FEBS Letters

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With the aging of population, neurological disorders, and especially disorders involving defects in protein conformation (also known as proteopathies) pose a serious socio‐economic problem. So far there is no effective treatment for most proteopathies, including Parkinson's disease (PD). The mechanism underlying PD pathogenesis is largely unknown, and the hallmark proteins, α‐synuclein (SYN) and tubulin polymerization promoting protein (TPPP/p25) are challenging drug targets. These proteins are intrinsically disordered with high conformational plasticity, and have diverse physiological and pathological functions. In the healthy brain, SYN and TPPP/p25 occur in neurons and oligodendrocytes, respectively; however, in PD and multiple system atrophy, they are co‐enriched and co‐localized in both cell types, thereby marking pathogenesis. Although large inclusions appear at a late disease stage, small, soluble assemblies of SYN promoted by TPPP/p25 are pathogenic. In the light of these issues, we established a new innovative strategy for the validation of a specific drug target based upon the identification of contact surfaces of the pathological SYN‐TPPP/p25 complex that may lead to the development of peptidomimetic foldamers suitable for pharmaceutical intervention.

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Targeting the interface of the pathological complex of α-synuclein and TPPP/p25

Szunyogh

Oláh

Szénási

et al. 2015

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Diseas

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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.

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Identification of motives mediating alternative functions of the neomorphic moonlighting TPPP/p25

Cited by 25 publications

References 35 publications

Challenging drug target for Parkinson's disease: Pathological complex of the chameleon TPPP/p25 and alpha-synuclein proteins

Challenging drug target for Parkinson's disease: Pathological complex of the chameleon TPPP/p25 and alpha-synuclein proteins

Pharmacological targeting of α‐synuclein and TPPP/p25 in Parkinson's disease: challenges and opportunities in a Nutshell

Targeting the interface of the pathological complex of α-synuclein and TPPP/p25

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