Polymer Nanoarray Approach for the Characterization of Biomolecular Interactions

Maity, Sibaprasad; Viazovkina, Ekaterina; Gall, A. A.; Lyubchenko, Yuri L.

doi:10.1007/978-1-4939-8591-3_5

Cited by 5 publications

(9 citation statements)

References 13 publications

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“…Figure 1A illustrates the FNA assembly of Aβ(14–23) monomers within the polymer, in which both monomers (shown as brown bars) are covalently attached at selected sites within the FNA polymer (shown in blue) using click chemistry (Figure S1). 28 The distance between the monomers in extended form is ~12 nm. Due to high FNA flexibility, the monomers are assembled into dimers as shown schematically on the right of Figure 1A.…”

Section: Resultsmentioning

confidence: 99%

“…The detailed characterization of trimers and tetramers was possible due to the use of our FNA approach to assemble dimers. 28, 41 This approach can be extended to oligomers longer than dimers and opens prospects for the characterization of larger order oligomers, which is our future goal.…”

Section: Resultsmentioning

confidence: 99%

“…28 HPLC purified cysteine-labeled HQKLVFFAED Aβ(14–23) was obtained from Peptide 2.0 (VA, USA). Cyanine 3-Maleimide (Cy3-MAL) was purchased from Lumiprobe Corporation (Florida, USA).…”

Section: Methodsmentioning

confidence: 99%

“…24–26 AFM force spectroscopy was used to probe interactions of Aβ(14–23) with its hairpin design. 27 These studies revealed that the interaction between the Aβ(14–23) monomer with the hairpin is stronger than the interaction between two monomers. These findings suggest that the hairpin structure contributes to oligomer stability.…”

Section: Introductionmentioning

confidence: 99%

“…One of these approaches is termed flexible nano array (FNA), in which the same two peptides are end-tethered inside of a flexible polymer to allow them to interact and form a dimer. 28 This idea has been implemented in this paper for probing the trimer and tetramer. The advantages of AFM force spectroscopy and TAPIN approaches are that they can be employed in a broad range of environmental conditions, such as ionic strength and pH.…”

Section: Introductionmentioning

confidence: 99%

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Single-molecule probing of amyloid nano-ensembles using the polymer nanoarray approach

Maity

Viazovkina

Gall

et al. 2017

Phys. Chem. Chem. Phys.

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Soluble amyloid-beta (Aβ) oligomers are the prime causitive agents of cognitive deficits during early stages of Alzheimer’s disease (AD). The transient nature of the oligomers makes them difficult to characterize by traditional techniques, suggesting that advanced approaches are necessary. Previously developed fluorescence-based tethered approach for probing intermolecular interactions (TAPIN) and AFM-based single-molecule force spectroscopy are capable of probing dimers of Aβ peptides. In this paper, a novel polymer nanoarray approach to probe trimers and tetramers formed by the Aβ(14–23) segment of Aβ protein at the single-molecule level is applied. By using this approach combined with TAPIN and AFM force spectroscopy, the impact of pH on the assembly of these oligomers was characterized. Experimental results reveal that pH affects the oligomer assembly process. At neutral pH, trimers and tetramers assemble into structures with a similar stability, while at acidic conditions (pH 3.7), the oligomers adopt a set of structures with different lifetimes and strengths. Models for the assembly of Aβ(14–23) trimers and tetramers based on the results obtained is proposed.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

“…28 HPLC purified cysteine-labeled HQKLVFFAED Aβ(14–23) was obtained from Peptide 2.0 (VA, USA). Cyanine 3-Maleimide (Cy3-MAL) was purchased from Lumiprobe Corporation (Florida, USA).…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Single-molecule probing of amyloid nano-ensembles using the polymer nanoarray approach

Maity

Viazovkina

Gall

et al. 2017

Phys. Chem. Chem. Phys.

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Probing Intermolecular Interactions within the Amyloid β Trimer Using a Tethered Polymer Nanoarray

2018

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Amyloid oligomers are considered the most neurotoxic species of amyloid aggregates. Spontaneous assembly of amyloids into aggregates is recognized as a major molecular mechanism behind Alzheimer's disease and other neurodegenerative disorders involving protein aggregation. Characterization of such oligomers is extremely challenging but complicated by their transient nature. Previously, we introduced a flexible nanoarray (FNA) method enabling us to probe dimers assembled by the amyloid β (14-23) [Aβ (14-23)] peptide. The study presented herein modifies and enhances this approach to assemble and probe trimers of Aβ (14-23). A metal-free click chemistry approach was used, in which dibenzocyclooctyne (DBCO) groups were incorporated at selected sites within the FNA template to click Aβ (14-23) monomers at their terminal azide groups. Atomic force microscopy (AFM) force spectroscopy was employed to characterize the assemblies. The force measurement data demonstrate that the dissociation of the trimer undergoes a stepwise pattern, in which the first monomer dissociates at the rupture force ∼48 ± 2.4 pN. The remaining dimer ruptures at the second step at a slightly larger rupture force (∼53 ± 3.2 pN). The assembled trimer was found to be quite dynamic, and transient species of this inherently dynamic process were identified.

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Design, Synthesis, and Properties of a Chemically Tethered Amyloid-β Segment Trimer Resistant to Intertrimer Mis-aggregation

2019

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The oligomer species of amyloid-β peptide (Aβ) may be relevant to the development of Alzheimer's disease. Isolating specific oligomer species of Aβ (i.e., dimers, trimers, tetramers, etc.), however, is difficult due to the transient, labile property of the oligomers. Here, we improved the resistance to intertrimer mis-aggregation of chemically tethered Aβ 25−35 trimers by introducing charged structures to the cyclic peptide tether that is covalently attached to the Aβ chain. The resistance to aggregation of the chemically tethered trimers positively correlated with the number of negative charges at the tether. Thus, a chemical trimer possessing three malonic acids at the tether exhibited high resistance because of the attenuated self-association by anionic repulsion. In addition, the malonic acid trimer possessed amyloidogenic properties such as cross-β-sheet structures, seeding activity, and cytotoxicity. This is the first study demonstrating that chemical modifications at the non-Aβ component enhance the resistance to aggregation of chemically tethered Aβ oligomers, by which the structural integrity of Aβ is maintained. Biological/biophysical evaluations of the intertrimer aggregation-resistant trimer may offer new, useful insights into the pathological functions of Aβ oligomers. Article pubs.acs.org/joc

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Polymer Nanoarray Approach for the Characterization of Biomolecular Interactions

Cited by 5 publications

References 13 publications

Single-molecule probing of amyloid nano-ensembles using the polymer nanoarray approach

Single-molecule probing of amyloid nano-ensembles using the polymer nanoarray approach

Probing Intermolecular Interactions within the Amyloid β Trimer Using a Tethered Polymer Nanoarray

Design, Synthesis, and Properties of a Chemically Tethered Amyloid-β Segment Trimer Resistant to Intertrimer Mis-aggregation

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