Peroxin-14,Peroxisomal membrane anchor protein PEX14,Peroxisomal membrane protein PEX14,Pex14,PTS1 receptor-docking protein,Rat,Rattus norvegicus
- Known as:
- Peroxin-14,Peroxisomal membrane anchor protein PEX14,Peroxisomal membrane protein PEX14,Pex14,PTS1 receptor-docking protein,Rat,Rattus norvegicus
- Catalog number:
- EIAAB30609
- Category:
- -
- Supplier:
- EIAab
- Gene target:
- Peroxin-14 Peroxisomal membrane anchor protein PEX14 Pex14 PTS1 receptor-docking Rat Rattus norvegicus
Related genes to: Peroxin-14,Peroxisomal membrane anchor protein PEX14,Peroxisomal membrane protein PEX14,Pex14,PTS1 receptor-docking protein,Rat,Rattus norvegicus
- Gene:
- PEX5 NIH gene
- Name:
- peroxisomal biogenesis factor 5
- Previous symbol:
- PXR1
- Synonyms:
- PTS1R
- Chromosome:
- 12p13.31
- Locus Type:
- gene with protein product
- Date approved:
- 1994-12-13
- Date modifiied:
- 2017-07-07
- Gene:
- PEX14 NIH gene
- Name:
- peroxisomal biogenesis factor 14
- Previous symbol:
- -
- Synonyms:
- -
- Chromosome:
- 1p36.22
- Locus Type:
- gene with protein product
- Date approved:
- 1998-08-21
- Date modifiied:
- 2014-11-18
Related products to: Peroxin-14,Peroxisomal membrane anchor protein PEX14,Peroxisomal membrane protein PEX14,Pex14,PTS1 receptor-docking protein,Rat,Rattus norvegicus
Related articles to: Peroxin-14,Peroxisomal membrane anchor protein PEX14,Peroxisomal membrane protein PEX14,Pex14,PTS1 receptor-docking protein,Rat,Rattus norvegicus
- Import of proteins into peroxisomes depends on PEX5, PEX13 and PEX14. By combining biochemical methods and structural biology, we show that the C-terminal SH3 domain of PEX13 mediates intramolecular interactions with a proximal FxxxF motif. The SH3 domain also binds WxxxF peptide motifs in the import receptor PEX5, demonstrating evolutionary conservation of such interactions from yeast to human. Strikingly, intramolecular interaction of the PEX13 FxxxF motif regulates binding of PEX5 WxxxF/Y motifs to the PEX13 SH3 domain. Crystal structures reveal how FxxxF and WxxxF/Y motifs are recognized by a non-canonical surface on the SH3 domain. The PEX13 FxxxF motif also mediates binding to PEX14. Surprisingly, the potential PxxP binding surface of the SH3 domain does not recognize PEX14 PxxP motifs, distinct from its yeast ortholog. Our data show that the dynamic network of PEX13 interactions with PEX5 and PEX14, mediated by diaromatic peptide motifs, modulates peroxisomal matrix import. - Source: PubMed
Publication date: 2024/04/18
Gaussmann StefanPeschel RebeccaOtt JuliaZak Krzysztof MSastre JuditDelhommel FlorentPopowicz Grzegorz MBoekhoven JobSchliebs WolfgangErdmann RalfSattler Michael - Protein-protein interactions (PPIs) constitute an important but challenging class of molecular targets for small molecules. The PEX5-PEX14 PPI has been shown to play a critical role in glycosome biogenesis and its disruption impairs the metabolism in Trpanosoma parasites, eventually leading to their death. Therefore, this PPI is a potential molecular target for new drugs against diseases caused by Trypanosoma infections. Here, we report a new class of peptidomimetic scaffolds to target the PEX5-PEX14 PPI. The molecular design was based on an oxopiperazine template for the α-helical mimetics. A structural simplification along with modifications of the central oxopiperazine scaffold and addressing the lipophilic interactions led to the development of peptidomimetics that inhibit PEX5-TbPEX14 PPI and display cellular activity against T. b. brucei. This approach provides an alternative approach towards the development of trypanocidal agents and may be generally useful for the design of helical mimetics as PPI inhibitors. - Source: PubMed
Publication date: 2023/06/25
Marciniak MonikaMróz PiotrNapolitano ValeriaKalel Vishal CFino RobertoPykacz EmiliaSchliebs WolfgangPlettenburg OliverErdmann RalfSattler MichaelPopowicz Grzegorz MDawidowski Maciej - Peroxisomes are organelles that carry out β-oxidation of fatty acids and amino acids. Both rare and prevalent diseases are caused by their dysfunction. Among disease-causing variant genes are those required for protein transport into peroxisomes. The peroxisomal protein import machinery, which also shares similarities with chloroplasts, is unique in transporting folded and large, up to 10 nm in diameter, protein complexes into peroxisomes. Current models postulate a large pore formed by transmembrane proteins; however, so far, no pore structure has been observed. In the budding yeast Saccharomyces cerevisiae, the minimum transport machinery includes the membrane proteins Pex13 and Pex14 and the cargo-protein-binding transport receptor, Pex5. Here we show that Pex13 undergoes liquid-liquid phase separation (LLPS) with Pex5-cargo. Intrinsically disordered regions in Pex13 and Pex5 resemble those found in nuclear pore complex proteins. Peroxisomal protein import depends on both the number and pattern of aromatic residues in these intrinsically disordered regions, consistent with their roles as 'stickers' in associative polymer models of LLPS. Finally, imaging fluorescence cross-correlation spectroscopy shows that cargo import correlates with transient focusing of GFP-Pex13 and GFP-Pex14 on the peroxisome membrane. Pex13 and Pex14 form foci in distinct time frames, suggesting that they may form channels at different saturating concentrations of Pex5-cargo. Our findings lead us to suggest a model in which LLPS of Pex5-cargo with Pex13 and Pex14 results in transient protein transport channels. - Source: PubMed
Publication date: 2023/05/10
Ravindran RiniBacellar Isabel O LCastellanos-Girouard XavierWahba Haytham MZhang ZhenghaoOmichinski James GKisley LydiaMichnick Stephen W - The cycling import receptor PEX5 and its membrane-located binding partner PEX14 are key constituents of the peroxisomal import machinery. Upon recognition of newly synthesized cargo proteins carrying a peroxisomal targeting signal type 1 (PTS1) in the cytosol, the PEX5/cargo complex docks at the peroxisomal membrane by binding to PEX14. The PEX14 N-terminal domain (NTD) recognizes (di)aromatic peptides, mostly corresponding to Wxxx(F/Y)-motifs, with nano-to micromolar affinity. Human PEX5 possesses eight of these conserved motifs distributed within its 320-residue disordered N-terminal region. Here, we combine biophysical (ITC, NMR, CD), biochemical and computational methods to characterize the recognition of these (di)aromatic peptides motifs and identify key features that are recognized by PEX14. Notably, the eight motifs present in human PEX5 exhibit distinct affinities and energetic contributions for the interaction with the PEX14 NTD. Computational docking and analysis of the interactions of the (di)aromatic motifs identify the specific amino acids features that stabilize a helical conformation of the peptide ligands and mediate interactions with PEX14 NTD. We propose a refined consensus motif ΦΦ for high affinity binding to the PEX14 NTD and discuss conservation of the (di)aromatic peptide recognition by PEX14 in other species. - Source: PubMed
Publication date: 2022/11/28
Gopalswamy MohanrajZheng ChenGaussmann StefanKooshapur HamedHambruch EvaSchliebs WolfgangErdmann RalfAntes IrisSattler Michael - The import of the majority of soluble peroxisomal proteins is initiated by the interaction between type-1 peroxisomal targeting signals (PTS1) and their receptor PEX5. PTS1 motifs reside at the extreme C-terminus of proteins and consist of a characteristic tripeptide and a modulatory upstream region. Various PTS1-PEX5 interactions have been studied by biophysical methods using isolated proteins or in heterologous systems such as two-hybrid assays, but a recently established approach based on Försters resonance energy transfer (FRET) allows a quantifying investigation in living cells. FRET is the radiation-free energy transfer between two fluorophores in close proximity and can be used to estimate the fraction of acceptor molecules bound to a donor molecule. For PTS1-PEX5 this method relies on the measurement of FRET-efficiency between the PTS1-binding TPR-domain of PEX5 tagged with mCherry and EGFP fused to a PTS1 peptide. However, this method is less suitable for binding partners with low affinity and protein complexes involving large proteins such as the interaction between full-length PTS1-carrying cargo proteins and PEX5. To overcome this limitation, we introduce a life-cell competition assay based on the same FRET approach but including a fusion protein of Cerulean with the protein of interest as a competitor. After implementing the mathematical description of competitive binding experiments into a fitting algorithm, we demonstrate the functionality of this approach using known interaction partners, its ability to circumvent previous limitations of FRET-measurements and its ability to study the interaction between PEX5 and its full-length cargo proteins. We find that some proteins (SCP2 and AGXT) bind PEX5 with higher affinity than their PTS1-peptides alone, but other proteins (ACOX3, DAO, PerCR-SRL) bind with lower but reasonable affinity, whereas GSTK1 binds with very low affinity. This binding strength was not increased upon elongating the PEX5 TPR-domain at its N-terminus, PEX5(N-TPR), although it interacts specifically with the N-terminal domain of PEX14. Finally, we demonstrate that the latter reduces the interaction strength between PEX5(N-TPR) and PTS1 by a dose-dependent but apparently non-competitive mechanism. Altogether, this demonstrates the power of this novel FRET-based competition approach for studying cargo recognition by PEX5 and protein complexes including large proteins in general. - Source: PubMed
Publication date: 2022/11/03
Hochreiter BernhardMalagon-Vina HugoSchmid Johannes ABerger JohannesKunze Markus