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Formation of Ordered Biomolecular Structures by the Self-assembly of Short Peptides

Published: November 21st, 2013



1Institute of Chemistry and The Center for Nanoscience and Nanotechnology, The Hebrew University of Jerusalem

This paper describes the formation of highly ordered peptide-based structures by the spontaneous process of self-assembly. The method utilizes commercially available peptides and common lab equipment. This technique can be applied to a large variety of peptides and may lead to the discovery of new peptide-based assemblies.

In nature, complex functional structures are formed by the self-assembly of biomolecules under mild conditions. Understanding the forces that control self-assembly and mimicking this process in vitro will bring about major advances in the areas of materials science and nanotechnology. Among the available biological building blocks, peptides have several advantages as they present substantial diversity, their synthesis in large scale is straightforward, and they can easily be modified with biological and chemical entities1,2. Several classes of designed peptides such as cyclic peptides, amphiphile peptides and peptide-conjugates self-assemble into ordered structures in solution. Homoaromatic dipeptides, are a class of short self-assembled peptides that contain all the molecular information needed to form ordered structures such as nanotubes, spheres and fibrils3-8. A large variety of these peptides is commercially available.

This paper presents a procedure that leads to the formation of ordered structures by the self-assembly of homoaromatic peptides. The protocol requires only commercial reagents and basic laboratory equipment. In addition, the paper describes some of the methods available for the characterization of peptide-based assemblies. These methods include electron and atomic force microscopy and Fourier-Transform Infrared Spectroscopy (FT-IR). Moreover, the manuscript demonstrates the blending of peptides (coassembly) and the formation of a "beads on a string"-like structure by this process.9 The protocols presented here can be adapted to other classes of peptides or biological building blocks and can potentially lead to the discovery of new peptide-based structures and to better control of their assembly.

Nature forms ordered and functional structures by the process of biomolecular self-assembly. Understanding the forces that govern this spontaneous process may lead to the ability to mimic self-assembly in vitro and consequently to major advances in the area of material sciences10,11. Peptides, specifically, hold great promise as a biomolecular building block, since they present large structural diversity, ease of chemical synthesis, and can easily be functionalized with biological and chemical entities. The field of peptide self-assembly was pioneered by Ghadiri and his colleagues, who demonstrated the self-assembly of peptide nanotubes by cyclic p....

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1. Self-assembly of Homoaromatic Dipeptides

  1. Weigh the desired peptide in its lyophilized form (e.g. NH2-Phe-Phe-OH, Boc-Phe-Phe-COOH) and prepare a stock solution by dissolving the peptide in 1,1,1,3,3,3-hexafluoro-2-propanol (HFP) to the appropriate concentration (e.g. 100 mg/ml for NH2-Phe-Phe-OH and Boc-Phe-Phe-COOH)7,17,46.
  2. Mix the solution using vortex and place on the bench until the peptide is completely dissolved and the solut.......

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This paper describes a method for the formation of ordered structures at the nano-and micrometer scale by the self-assembly of peptides. In order to demonstrate this simple process we present and characterize the coassembly of two simple aromatic peptides (Figure 1). One of the peptides is the NH2-Phe-Phe-OH (diphenylalanine) peptide, which can self-assemble in an aqueous solution into hollow tubular structures with nanometric dimensions7. The other peptide is its Boc protected anal.......

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In summary, this paper demonstrates the ease in which peptide-based assemblies can be formed in vitro. The process involves commercially available peptides and solvents, and it occurs spontaneously under ambient conditions, upon the addition of a polar solvent to the test tube. It is crucial to use HFP as a solvent of the peptides, due to the low solubility of the peptides in other organic solvents. In addition, due to the high volatility of HFP it is necessary to prepare fresh stock solution for each experiment.......

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This work was supported by the Marie Curie International Reintegration Grant and by the German-Israel Foundation. We acknowledge Mr. Yair Razvag for AFM analysis.


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NameCompanyCatalog NumberComments
Name of Material/ EquipmentCompanyCatalog NumberComments/Description
Ethanol absolute (Dehydrated) AR sterileBio-Lab Ltd.52555Blending with TDW for the preparation of 50% solution
Uranyl acetateSigma-Aldrich73943For negative staining. It is possible to work without it.
glass cover slipMarienfeld Laboratory Glassware110590
TEM gridsElectron Microscopy SciencesFCF200-Cu-50Formvar/Carbon 200 Mesh, Cu
Quantitive filter paperWhatman1001055
Deuterium Oxide (D2O)Sigma-Aldrich151882-100G99.9 atom % D
CaF2 windowPIKE Technologies160-121225 mm x 2 mm window. For FT-IR measurments
AFM tipsNanoScience InstrumentsCFMRAspire probes, CFMR-25 series
Filter unitsMilliporeSLGV033RSMillex-GV, 0.22 μm, PVDF, 33 mm, gamma sterilized
TEMFEITecnai T12 G2 Spirit
AFMJPK InstrumentsA JPK NanoWizard3
FT-IRThermo Fisher ScientificNicolet 6700 advanced gold spectrometer
FT-IR PurgeParkerBALSTON FT-IR Purge Gas Generator model 75-52
OMNIC (Nicolet) softwareThermo Nicolet CorporationFor FT-IR spectra analysis
Vortex mixerWisd Laboratory EquipmentViseMix VM
WeightMettler ToledoNewClassic MS
Sputter coaterPolaronSC7640 Sputter Coater

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