Probing Conformational Change of Intrinsically Disordered α-Synuclein to Helical Structures by Distinctive Regional Interactions with Lipid Membranes

2014 ◽  
Vol 86 (3) ◽  
pp. 1909-1916 ◽  
Author(s):  
Shin Jung C. Lee ◽  
Jong Wha Lee ◽  
Tae Su Choi ◽  
Kyeong Sik Jin ◽  
Seonghwan Lee ◽  
...  
Keyword(s):  
Conformational Change ◽  
Lipid Membranes ◽  
Helical Structures ◽  
Intrinsically Disordered

scholarly journals Yeast surface display of full-length human microtubule-associated protein tau

2019 ◽  
Author(s):  
Shiyao Wang ◽  
Yong Ku Cho
Keyword(s):  
Conformational Change ◽  
Protein Interactions ◽  
Surface Display ◽  
Full Length ◽  
Adult Brain ◽  
Yeast Surface Display ◽  
Post Translational Modification ◽  
Protein Tau ◽  
Microtubule Associated Protein Tau ◽  
Intrinsically Disordered

AbstractMicrotubule-associated protein tau is an intrinsically-disordered, highly soluble protein found primarily in neurons. Under normal conditions, tau regulates the stability of axonal microtubules and intracellular vesicle transport. However, in patients of neurodegeneration such as Alzheimer’s disease (AD), tau forms neurofibrillary deposits, which correlates well with the disease progression. Identifying molecular signatures in tau, such as post-translational modification, truncation, and conformational change has great potential to detect earliest signs of neurodegeneration, and develop therapeutic strategies. Here we show that full-length human tau, including the longest isoform found in the adult brain can be robustly displayed on the surface of yeastSaccharomyces cerevisiae. Yeast-displayed tau binds to anti-tau antibodies that cover epitopes ranging from the N-terminus to the 4R repeat region. Unlike tau expressed in the yeast cytosol, surface-displayed tau was not phosphorylated at sites found in AD patients (probed by antibodies AT8, AT270, AT180, PHF-1). However, yeast-displayed tau showed clear binding to paired helical filament (PHF) tau conformation-specific antibodies Alz-50, MC-1, and Tau-2. Although the tau possessed a conformation found in PHFs, oligomerization or aggregation into larger filaments were undetected. Taken together, yeast-displayed tau enables robust measurement of protein interactions, and is of particular interest for characterizing conformational change.

scholarly journals Living in Promiscuity: The Multiple Partners of Alpha-Synuclein at the Synapse in Physiology and Pathology

2019 ◽  
Vol 20 (1) ◽  
pp. 141 ◽  
Author(s):  
Francesca Longhena ◽  
Gaia Faustini ◽  
Maria Grazia Spillantini ◽  
Arianna Bellucci
Keyword(s):  
Lipid Membranes ◽  
Intrinsically Disordered Proteins ◽  
Lipid Membrane ◽  
Lewy Bodies ◽  
Alpha Synuclein ◽  
Disordered Proteins ◽  
Monoamine Transporters ◽  
Post Translational Modification ◽  
Intrinsically Disordered ◽  
Multiple Partners

Alpha-synuclein (α-syn) is a small protein that, in neurons, localizes predominantly to presynaptic terminals. Due to elevated conformational plasticity, which can be affected by environmental factors, in addition to undergoing disorder-to-order transition upon interaction with different interactants, α-syn is counted among the intrinsically disordered proteins (IDPs) family. As with many other IDPs, α-syn is considered a hub protein. This function is particularly relevant at synaptic sites, where α-syn is abundant and interacts with many partners, such as monoamine transporters, cytoskeletal components, lipid membranes, chaperones and synaptic vesicles (SV)-associated proteins. These protein–protein and protein–lipid membrane interactions are crucial for synaptic functional homeostasis, and alterations in α-syn can cause disruption of this complex network, and thus a failure of the synaptic machinery. Alterations of the synaptic environment or post-translational modification of α-syn can induce its misfolding, resulting in the formation of oligomers or fibrillary aggregates. These α-syn species are thought to play a pathological role in neurodegenerative disorders with α-syn deposits such as Parkinson’s disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA), which are referred to as synucleinopathies. Here, we aim at revising the complex and promiscuous role of α-syn at synaptic terminals in order to decipher whether α-syn molecular interactants may influence its conformational state, contributing to its aggregation, or whether they are just affected by it.

scholarly journals Interaction of two intrinsically disordered plant stress proteins (COR15A and COR15B) with lipid membranes in the dry state

2010 ◽  
Vol 1798 (9) ◽  
pp. 1812-1820 ◽  
Author(s):  
Anja Thalhammer ◽  
Michaela Hundertmark ◽  
Antoaneta V. Popova ◽  
Robert Seckler ◽  
Dirk K. Hincha
Keyword(s):  
Lipid Membranes ◽  
Stress Proteins ◽  
Plant Stress ◽  
Intrinsically Disordered

Pro-apoptotic bax-α1 synthesis and evidence for β-sheet to α-helix conformational change as triggered by negatively charged lipid membranes

2007 ◽  
Vol 13 (2) ◽  
pp. 100-106 ◽  
Author(s):  
Marc-Antoine Sani ◽  
Cécile Loudet ◽  
Gerhard Gröbner ◽  
Erick J. Dufourc
Keyword(s):  
Conformational Change ◽  
Lipid Membranes ◽  
Α Helix ◽  
Β Sheet

Dissecting mechanism of coupled folding and binding of an intrinsically disordered protein by chemical synthesis of conformationally constrained analogues

2017 ◽  
Vol 53 (53) ◽  
pp. 7369-7372 ◽  
Author(s):  
Boris Schmidtgall ◽  
Olivier Chaloin ◽  
Valentin Bauer ◽  
Manuela Sumyk ◽  
Catherine Birck ◽  
...  
Keyword(s):  
Amino Acids ◽  
Chemical Synthesis ◽  
Intrinsically Disordered Protein ◽  
Helical Structures ◽  
Activation Domain ◽  
Conformationally Constrained ◽  
Intrinsically Disordered ◽  
Disordered Protein

Non-canonical α-methyl amino acids were incorporated at various sites in the sequence of intrinsically disordered activation domain from the p160 transcriptional co-activator (ACTR) to facilitate the formation of α-helical structures.

scholarly journals Nucleic Acid-Dependent Structural Transition of the Intrinsically Disordered N-Terminal Appended Domain of Human Lysyl-tRNA Synthetase

2018 ◽  
Vol 19 (10) ◽  
pp. 3016 ◽  
Author(s):  
Soon Kwon ◽  
Ji Yu ◽  
Chan Park ◽  
Jiseop Lee ◽  
Baik Seong
Keyword(s):  
Structural Transition ◽  
Rna Binding ◽  
Functional Organization ◽  
Affinity Purification ◽  
Trna Synthetase ◽  
Helical Conformation ◽  
Helical Structures ◽  
Interaction Domain ◽  
Intrinsically Disordered

Eukaryotic lysyl-tRNA synthetases (LysRS) have an N-terminal appended tRNA-interaction domain (RID) that is absent in their prokaryotic counterparts. This domain is intrinsically disordered and lacks stable structures. The disorder-to-order transition is induced by tRNA binding and has implications on folding and subsequent assembly into multi-tRNA synthetase complexes. Here, we expressed and purified RID from human LysRS (hRID) in Escherichia coli and performed a detailed mutagenesis of the appended domain. hRID was co-purified with nucleic acids during Ni-affinity purification, and cumulative mutations on critical amino acid residues abolished RNA binding. Furthermore, we identified a structural ensemble between disordered and helical structures in non-RNA-binding mutants and an equilibrium shift for wild-type into the helical conformation upon RNA binding. Since mutations that disrupted RNA binding led to an increase in non-functional soluble aggregates, a stabilized RNA-mediated structural transition of the N-terminal appended domain may have implications on the functional organization of human LysRS and multi-tRNA synthetase complexes in vivo.

scholarly journals The intracellular lipid-binding domain of human Na+/H+ exchanger 1 forms a lipid-protein co-structure essential for activity

2020 ◽  
Vol 3 (1) ◽  
Author(s):  
Ruth Hendus-Altenburger ◽  
Jens Vogensen ◽  
Emilie Skotte Pedersen ◽  
Alessandra Luchini ◽  
Raul Araya-Secchi ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Lipid Membranes ◽  
Molecular Mechanisms ◽  
Lipid Binding ◽  
Proximal Region ◽  
Dynamic Interactions ◽  
Intrinsically Disordered ◽  
Nhe1 Activity ◽  
Dynamics Simulations ◽  
Membrane Anchoring

AbstractDynamic interactions of proteins with lipid membranes are essential regulatory events in biology, but remain rudimentarily understood and particularly overlooked in membrane proteins. The ubiquitously expressed membrane protein Na+/H+-exchanger 1 (NHE1) regulates intracellular pH (pHi) with dysregulation linked to e.g. cancer and cardiovascular diseases. NHE1 has a long, regulatory cytosolic domain carrying a membrane-proximal region described as a lipid-interacting domain (LID), yet, the LID structure and underlying molecular mechanisms are unknown. Here we decompose these, combining structural and biophysical methods, molecular dynamics simulations, cellular biotinylation- and immunofluorescence analysis and exchanger activity assays. We find that the NHE1-LID is intrinsically disordered and, in presence of membrane mimetics, forms a helical αα-hairpin co-structure with the membrane, anchoring the regulatory domain vis-a-vis the transport domain. This co-structure is fundamental for NHE1 activity, as its disintegration reduced steady-state pHi and the rate of pHi recovery after acid loading. We propose that regulatory lipid-protein co-structures may play equally important roles in other membrane proteins.

scholarly journals Ubiquitin Interacting Motifs: Duality Between Structured and Disordered Motifs

2021 ◽  
Vol 8 ◽  
Author(s):  
Matteo Lambrughi ◽  
Emiliano Maiani ◽  
Burcu Aykac Fas ◽  
Gary S. Shaw ◽  
Birthe B. Kragelund ◽  
...  
Keyword(s):  
Protein Function ◽  
Chemical Shifts ◽  
Helical Structures ◽  
Conserved Sequence ◽  
Intrinsically Disordered ◽  
Intrinsically Disordered Regions ◽  
Multiple Partners ◽  
Common Motif ◽  
Dynamics Simulations ◽  
Disordered Regions

Ubiquitin is a small protein at the heart of many cellular processes, and several different protein domains are known to recognize and bind ubiquitin. A common motif for interaction with ubiquitin is the Ubiquitin Interacting Motif (UIM), characterized by a conserved sequence signature and often found in multi-domain proteins. Multi-domain proteins with intrinsically disordered regions mediate interactions with multiple partners, orchestrating diverse pathways. Short linear motifs for binding are often embedded in these disordered regions and play crucial roles in modulating protein function. In this work, we investigated the structural propensities of UIMs using molecular dynamics simulations and NMR chemical shifts. Despite the structural portrait depicted by X-crystallography of stable helical structures, we show that UIMs feature both helical and intrinsically disordered conformations. Our results shed light on a new class of disordered UIMs. This group is here exemplified by the C-terminal domain of one isoform of ataxin-3 and a group of ubiquitin-specific proteases. Intriguingly, UIMs not only bind ubiquitin. They can be a recruitment point for other interactors, such as parkin and the heat shock protein Hsc70-4. Disordered UIMs can provide versatility and new functions to the client proteins, opening new directions for research on their interactome.

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