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Structural analysis of the neuropeptide substance P by using vibrational spectroscopy

  • Zuzana JurasekovaEmail author
  • Adianez Garcia-Leis
  • Santiago Sanchez-Cortes
  • Anna Tinti
  • Armida TorreggianiEmail author
Research Paper
  • 36 Downloads

Abstract

Substance P (SP) is one of the most studied peptide hormones and knowing the relationship between its structure and function may have important therapeutic applications in the treatment of a variety of stress-related illnesses. In order to obtain a deeper insight into its folding, the effects of different factors, such as pH changes, the presence of Ca2+ ions, and the substitution of the Met-NH2 moiety in the SP structure, was studied by Raman and infrared spectroscopies. SP has a pH-dependent structure. Under acidic–neutral conditions, SP possesses a prevalent β-sheet structure although also other secondary structure elements are present. By increasing pH, a higher orderliness in the SP secondary structure is induced, as well as the formation of strongly bound intermolecular β-strands with a parallel alignment, which favour the self-assembly of SP in β-aggregates. The substitution of the Met-NH2 moiety with the acidic functional group in the SP sequence, giving rise to a not biologically active SP analogue, results in a more disordered folding, where the predominant contribution comes from a random coil. Conversely, the presence of Ca2+ ions affects slightly but sensitively the folding of the polypeptide chain, by favouring the α-helical content and a different alignment of β-strands; these are structural elements, which may favour the SP biological activity. In addition, the capability of SERS spectroscopy to detect SP in its biologically active form was also tested by using different metal nanoparticles. Thanks to the use of silver NPs prepared by reduction of silver nitrate with hydroxylamine hydrochloride, SP can be detected at very low peptide concentration (~ 90 nM). However, the SERS spectra cannot be obtained under alkaline conditions since both the formation of SP aggregates and the lack of ion pairs do not allow a strong enough interaction of SP with silver NPs.

Graphical abstract

Keywords

Substance P Vibrational (Raman and infrared) spectroscopy Surface-enhanced Raman spectroscopy (SERS) 

Abbreviations

(Ag/Au) NPs

(Silver/gold) Nanoparticles

FWHM

Full width at half maximum

LOD

Limit of detection

SP

Substance P in amidated form

SP-OH

Substance P in acidic form

SERS

Surface-enhanced Raman scattering

Arg

Arginine

Gly

Glycine

Ile

Isoleucine

Leu

Leucine

Lys

Lysine

Met

Methionine

Phe

Phenylalanine

Pro

Proline

Notes

Funding information

This work was supported by the Scientific Grant Agency of the Ministry of the Education of Slovak Republic (APVV-15-0485) and the Ministerio de Economía y Competitividad from Spain under the grant FIS2017-84314-R.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Supplementary material

216_2019_2097_MOESM1_ESM.pdf (692 kb)
ESM 1 (PDF 691 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Zuzana Jurasekova
    • 1
    • 2
    Email author
  • Adianez Garcia-Leis
    • 3
  • Santiago Sanchez-Cortes
    • 3
  • Anna Tinti
    • 4
  • Armida Torreggiani
    • 5
    Email author
  1. 1.Department of Biophysics, Faculty of ScienceP. J. Šafárik University in KošiceKošiceSlovakia
  2. 2.Center for Interdisciplinary Biosciences, Technology and Innovation ParkP. J. Šafárik University in KošiceKošiceSlovakia
  3. 3.Instituto de Estructura de la Materia, CSICMadridSpain
  4. 4.Dipartimento di Scienze Biomediche e NeuromotorieUniversità di BolognaBolognaItaly
  5. 5.Istituto per la Sintesi Organica e la FotoreattivitàConsiglio Nazionale delle Ricerche, CNRBolognaItaly

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