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Analytical and Bioanalytical Chemistry

, Volume 407, Issue 14, pp 4015–4021 | Cite as

Quantifying amyloid fibrils in protein mixtures via infrared attenuated-total-reflection spectroscopy

  • Pei Wang
  • Wilhelm Bohr
  • Markus Otto
  • Karin M. Danzer
  • Boris MizaikoffEmail author
Research Paper
Part of the following topical collections:
  1. Direct Optical Detection

Abstract

Amyloid aggregation of proteins is usually associated with amyloid diseases. A distinct feature of protein aggregation is the increase of crossed β-sheet structures. Infrared attenuated-total-reflectance (IR-ATR) spectroscopy is a sensitive optical technique that has the potential to provide secondary structure characteristics of proteins even in complex biological samples. In this study we report the analysis of secondary structures of proteins, using the amide I band for the detection and quantification of amyloid fibrils in protein mixtures by use of IR-ATR techniques, at comparatively low sample concentrations. From the experimental results, an analytical model of the relationship between the IR spectra of protein mixtures and the individual mixture components was established using spectral-deconvolution procedures and curve-fitting methods. On the basis of this model, four ratios were shown to provide direct information on amyloid aggregated fibrils via the increase of crossed β-sheet structures in protein-mixture samples. In conclusion, this study confirms the utility of IR spectroscopy for analyzing protein mixtures and for identifying amyloid fibril information within such complex multi-component samples.

Keywords

Amyloid fibrils Protein-mixture analysis Mid-infrared Spectral deconvolution Fourier-transform infrared spectroscopy (FTIR) Attenuated total reflection (ATR) 

Notes

Acknowledgments

Assistance of A. Wilk and X. Wang (Institute for Analytical and Bioanalytical Chemistry, Univ. Ulm) during initial IR-ATR studies is greatly acknowledged. We are grateful for a graduate student fellowship provided by the International Graduate School in Molecular Medicine at the University of Ulm. This study was in part supported by the BMBF (Federal Ministry of Education and Research, Germany), the Competence Net Neurodegenerative Dementias (project: FTLDc), the JNPD Networks for Standardization of Biomarkers (BiomarkAPD, Sophia), the EU (project: NADINE), the Foundation of the State of Baden-Württemberg, the Boehringer Ingelheim Ulm University BioCenter (BIU), and the State of Baden-Württemberg Junior Professor Programme.

References

  1. 1.
    Eisenberg D, Jucker M (2012) The amyloid state of proteins in human diseases. Cell 148:1188–1203CrossRefGoogle Scholar
  2. 2.
    Sunde M, Serpell LC, Bartlam M, Fraser PE, Pepys MB, Blake CC (1997) Common core structure of amyloid fibrils by synchrotron X-ray diffraction. J Mol Biol 273:729–739CrossRefGoogle Scholar
  3. 3.
    Jackson M, Mantsch HH (1995) The use and misuse of FTIR spectroscopy in the determination of protein structure. Crit Rev Biochem Mol Biol 30:95–120CrossRefGoogle Scholar
  4. 4.
    Mantsch HH, Chapman D (1996) Infrared spectroscopy of biomolecules. Wiley-Liss, New YorkGoogle Scholar
  5. 5.
    Barth A (2007) Infrared spectroscopy of proteins. Biochim Biophys Acta Bioenerg 1767:1073–1101CrossRefGoogle Scholar
  6. 6.
    Goormaghtigh E, Ruysschaert J-M, Raussens V (2006) Evaluation of the information content in infrared spectra for protein secondary structure determination. Biophys J 90:2946–2957CrossRefGoogle Scholar
  7. 7.
    Sarroukh R, Goormaghtigh E, Ruysschaert J-M, Raussens V (2013) ATR-FTIR: a “rejuvenated” tool to investigate amyloid proteins. BBA - Biomembr 1–11Google Scholar
  8. 8.
    Shivu B, Seshadri S, Li J, Oberg KA, Uversky VN, Fink AL (2013) Distinct β-sheet structure in protein aggregates determined by ATR–FTIR Spectroscopy. Biochemistry 52:5176–5183CrossRefGoogle Scholar
  9. 9.
    Hong D-P, Fink AL, Uversky VN (2008) Structural characteristics of α-synuclein oligomers stabilized by the Flavonoid Baicalein. J Mol Biol 383:214–223CrossRefGoogle Scholar
  10. 10.
    Fändrich M, Forge V, Buder K, Kittler M, Dobson CM, Diekmann S (2003) Myoglobin forms amyloid fibrils by association of unfolded polypeptide segments. Proc Natl Acad Sci U S A 100:15463–15468CrossRefGoogle Scholar
  11. 11.
    Griebe M, Daffertshofer M, Stroick M, Syren M, Ahmad-Nejad P, Neumaier M, Backhaus J, Hennerici MG, Fatar M (2007) Infrared spectroscopy: a new diagnostic tool in Alzheimer disease. Neurosci Lett 420:29–33CrossRefGoogle Scholar
  12. 12.
    Ahmed SSSJ, Santosh W, Kumar S, Thanka Christlet TH (2010) Neural network algorithm for the early detection of Parkinson's disease from blood plasma by FTIR micro-spectroscopy. Vib Spectrosc 53:181–188CrossRefGoogle Scholar
  13. 13.
    Yun M, Wu W, Hood L, Harrington M (1992) Human cerebrospinal fluid protein database: edition 1992. Electrophoresis 13:1002–1013CrossRefGoogle Scholar
  14. 14.
    Hühmer AF, Biringer RG, Amato H, Fonteh AN, Harrington MG (2006) Protein analysis in human cerebrospinal fluid: physiological aspects, current progress and future challenges. Dis Markers 22:3–26CrossRefGoogle Scholar
  15. 15.
    Fändrich M, Fletcher MA, Dobson CM (2001) Amyloid fibrils from muscle myoglobin. Nature 410:165–166CrossRefGoogle Scholar
  16. 16.
    Yan Y-B, Wang Q, He H-W, Hu X-Y, Zhang R-Q, Zhou H-M (2003) Two-dimensional infrared correlation spectroscopy study of sequential events in the heat-induced unfolding and aggregation process of myoglobin. Biophys J 85:1959–1967CrossRefGoogle Scholar
  17. 17.
    van de Weert M, Haris PI, Hennink WE, Crommelin DJ (2001) Fourier transform infrared spectrometric analysis of protein conformation: effect of sampling method and stress factors. Anal Biochem 297:160–169CrossRefGoogle Scholar
  18. 18.
    Fändrich M (2012) Oligomeric intermediates in amyloid formation: structure determination and mechanisms of toxicity. J Mol Biol 421:427–440CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Pei Wang
    • 1
  • Wilhelm Bohr
    • 1
  • Markus Otto
    • 2
  • Karin M. Danzer
    • 2
  • Boris Mizaikoff
    • 1
    Email author
  1. 1.Institute for Analytical and Bioanalytical ChemistryUlm UniversityUlmGermany
  2. 2.Department of NeurologyUlm UniversityUlmGermany

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