A chemometric approach for characterization of serum transthyretin in familial amyloidotic polyneuropathy type I (FAP-I) by electrospray ionization-ion mobility mass spectrometry

Talanta. 2018 May 1:181:87-94. doi: 10.1016/j.talanta.2017.12.072. Epub 2017 Dec 30.

Abstract

In this study, we describe a chemometric data analysis approach to assist in the interpretation of the complex datasets from the analysis of high-molecular mass oligomeric proteins by ion mobility mass spectrometry (IM-MS). The homotetrameric protein transthyretin (TTR) is involved in familial amyloidotic polyneuropathy type I (FAP-I). FAP-I is associated with a specific TTR mutant variant (TTR(Met30)) that can be easily detected analyzing the monomeric forms of the mutant protein. However, the mechanism of protein misfolding and aggregation onset, which could be triggered by structural changes in the native tetrameric protein, remains under investigation. Serum TTR from healthy controls and FAP-I patients was purified under non-denaturing conditions by conventional immunoprecipitation in solution and analyzed by IM-MS. IM-MS allowed separation and characterization of several tetrameric, trimeric and dimeric TTR gas ions due to their differential drift time. After an appropriate data pre-processing, multivariate curve resolution alternating least squares (MCR-ALS) was applied to the complex datasets. A group of seven independent components being characterized by their ion mobility profiles and mass spectra were resolved to explain the observed data variance in control and patient samples. Then, principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were considered for exploration and classification. Only four out of the seven resolved components were enough for an accurate differentiation. Furthermore, the specific TTR ions identified in the mass spectra of these components and the resolved ion mobility profiles provided a straightforward insight into the most relevant oligomeric TTR proteoforms for the disease.

Keywords: Curve resolution; Familial amyloidotic polyneuropathy; Ion mobility; Mass spectrometry; Multivariate data analysis; Neurodegeneration.

MeSH terms

  • Amyloid Neuropathies, Familial / blood*
  • Amyloid Neuropathies, Familial / genetics
  • Humans
  • Mutant Proteins / blood*
  • Mutant Proteins / chemistry
  • Mutant Proteins / isolation & purification
  • Prealbumin / analysis*
  • Prealbumin / chemistry
  • Prealbumin / genetics
  • Protein Multimerization
  • Proteomics / methods
  • Reproducibility of Results
  • Spectrometry, Mass, Electrospray Ionization / methods*

Substances

  • Mutant Proteins
  • Prealbumin