Unravelling the role of amino acid sequence order in the assembly and function of the amyloid-β core

Background: RADA-4 (Ac-RADARADARADARADA-NH2) is the most extensively studied and marketed self-assembling peptide, forming hydrogel, used to create defined threedimensional microenvironments for cell culture applications. Objectives: In this work, we use various biophysical techniques to investigate the length dependency of RADA aggregation and assembly. Methods: We synthesized a series of RADA-N peptides, N ranging from 1 to 4, resulting in four peptides having 4, 8, 12, and 16 amino acids in their sequence. Through a combination of various biophysical methods including thioflavin T fluorescence assay, static right angle light scattering assay, Dynamic Light Scattering (DLS), electron microscopy, CD, and IR spectroscopy, we have examined the role of chain-length on the self-assembly of RADA peptide. Results: Our observations show that the aggregation of ionic, charge-complementary RADA motifcontaining peptides is length-dependent, with N less than 3 are not forming spontaneous selfassemblies. Conclusion: The six biophysical experiments discussed in this paper validate the significance of chain-length on the epitaxial growth of RADA peptide self-assembly.

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Self-assembling behaviour of a modified aromatic amino acid in competitive medium

Soft Matter ◽

10.1039/d0sm00584c ◽

2020 ◽

Vol 16 (28) ◽

pp. 6599-6607 ◽

Cited By ~ 1

Author(s):

Pijush Singh ◽

Souvik Misra ◽

Nayim Sepay ◽

Sanjoy Mondal ◽

Debes Ray ◽

...

Keyword(s):

Amino Acid ◽

Self Assembly ◽

Aromatic Amino Acid ◽

Photophysical Properties ◽

Solvent Mixture ◽

The Self ◽

Solvent Ratio ◽

Self Assembling

The self-assembly and photophysical properties of 4-nitrophenylalanine (4NP) are changed with the alteration of solvent and final self-assembly state of 4NP in competitive solvent mixture and are dictated by the solvent ratio.

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Understanding the role of conjugation length on the self-assembly behaviour of oligophenyleneethynylenes

Chemical Communications ◽

10.1039/d1cc01054a ◽

2021 ◽

Author(s):

Beatriz Matarranz ◽

Goutam Ghosh ◽

Ramesh Kandanelli ◽

Angel Sampedro ◽

Kalathil K. Kartha ◽

...

Keyword(s):

Self Assembly ◽

The Self ◽

Conjugation Length ◽

The Relationship

We unravel the relationship between conjugation length and self-assembly behaviour of oligophenyleneethynylenes (OPEs).

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An evaluation of the self-assembly enhancing properties of cell-derived hexameric amyloid-β

Scientific Reports ◽

10.1038/s41598-021-90680-y ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Devkee M. Vadukul ◽

Céline Vrancx ◽

Pierre Burguet ◽

Sabrina Contino ◽

Nuria Suelves ◽

...

Keyword(s):

Amyloid Precursor Protein ◽

Self Assembly ◽

Critical Nucleus ◽

Amyloid Fibril ◽

Amyloid Β ◽

Amyloid Plaques ◽

The Self ◽

Aβ Peptide ◽

Amyloid Fibril Formation ◽

Secretase Activity

AbstractA key hallmark of Alzheimer’s disease is the extracellular deposition of amyloid plaques composed primarily of the amyloidogenic amyloid-β (Aβ) peptide. The Aβ peptide is a product of sequential cleavage of the Amyloid Precursor Protein, the first step of which gives rise to a C-terminal Fragment (C99). Cleavage of C99 by γ-secretase activity releases Aβ of several lengths and the Aβ42 isoform in particular has been identified as being neurotoxic. The misfolding of Aβ leads to subsequent amyloid fibril formation by nucleated polymerisation. This requires an initial and critical nucleus for self-assembly. Here, we identify and characterise the composition and self-assembly properties of cell-derived hexameric Aβ42 and show its assembly enhancing properties which are dependent on the Aβ monomer availability. Identification of nucleating assemblies that contribute to self-assembly in this way may serve as therapeutic targets to prevent the formation of toxic oligomers.

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Chitosanase from Streptomyces sp. strain N174: a comparative review of its structure and function

Biochemistry and Cell Biology ◽

10.1139/o97-079 ◽

1997 ◽

Vol 75 (6) ◽

pp. 687-696 ◽

Cited By ~ 20

Author(s):

Tamo Fukamizo ◽

Ryszard Brzezinski

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Substrate Binding ◽

Structure And Function ◽

Site Directed Mutagenesis ◽

Directed Mutagenesis ◽

Amino Acid Residues ◽

Streptomyces Sp ◽

Binding Cleft ◽

And Function

Novel information on the structure and function of chitosanase, which hydrolyzes the beta -1,4-glycosidic linkage of chitosan, has accumulated in recent years. The cloning of the chitosanase gene from Streptomyces sp. strain N174 and the establishment of an efficient expression system using Streptomyces lividans TK24 have contributed to these advances. Amino acid sequence comparisons of the chitosanases that have been sequenced to date revealed a significant homology in the N-terminal module. From energy minimization based on the X-ray crystal structure of Streptomyces sp. strain N174 chitosanase, the substrate binding cleft of this enzyme was estimated to be composed of six monosaccharide binding subsites. The hydrolytic reaction takes place at the center of the binding cleft with an inverting mechanism. Site-directed mutagenesis of the carboxylic amino acid residues that are conserved revealed that Glu-22 and Asp-40 are the catalytic residues. The tryptophan residues in the chitosanase do not participate directly in the substrate binding but stabilize the protein structure by interacting with hydrophobic and carboxylic side chains of the other amino acid residues. Structural and functional similarities were found between chitosanase, barley chitinase, bacteriophage T4 lysozyme, and goose egg white lysozyme, even though these proteins share no sequence similarities. This information can be helpful for the design of new chitinolytic enzymes that can be applied to carbohydrate engineering, biological control of phytopathogens, and other fields including chitinous polysaccharide degradation. Key words: chitosanase, amino acid sequence, overexpression system, reaction mechanism, site-directed mutagenesis.

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