Biopolym. Cell. 2014; 30(3):190-196.
http://dx.doi.org/10.7124/bc.000895
Structure and Function of Biopolymers
Application of MALDI-TOF mass spectrometry for study on fibrillar and oligomeric aggregates of alpha-synuclein
1Severinovskaya O. V., 2Kovalska V. B., 2Losytskyy M. Yu., 3Cherepanov V. V., 4Subramaniam V., 2Yarmoluk S. M.
  1. Chuyko Institute of Surface Chemistry, NAS of Ukraine
    17, Generala Naumova Str., Kyiv, Ukraine, 03164
  2. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
  3. Institute of Physics, NAS of Ukraine
    46, Prospect Nauki, Kyiv, Ukraine, 03028
  4. Nanobiophysics Group, MESA and Institute for Nanotechnology and MIRA Institute for Biomedical Technology, University of Twente
    PO Box 217, 7500 AE Enschede, The Netherlands

Abstract

Aim. To study the -synuclein (ASN) aggregates of different structural origin, namely amyloid fibrils and spherical oligomers, in comparison with a native protein. Methods. MALDI TOF mass spectrometry and atomic for- ce microscopy (AFM). Results. The mass spectra of native and fibrillar ASN have similar character, i. e. they are characterized by the well pronounced peak of protein molecular ion, the low molecular weight associates, and rather low contain of fragmentation products. The spectrum of oligomeric aggregate is characterized by the high contain of fragmentation products, low intensity of protein molecular ion and the absence of peaks of associates. Conclusions. The difference between the spectra of fibrillar and oligomeric ASN could be explained, first, by the different content of the «residual» monomeric ASN and the protein degradation products in the studied samples, and, second, by the different structure-depended mechanisms of the protein degradation induced by the laser ionization. We suggested that the MALDI-TOF mass spectroscopy is a method useful for the investigation of ASN aggregation and characterization of its high order self-associates; besides, there is an interest in estimating the potency of the MALDI-TOF for the analysis of aggregation of various amyloidogenic proteins.
Keywords: alpha-synuclein, MALDI-TOF, amyloid fibril, oligomeric aggregate, AFM
Full text: (PDF, in English)

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