Application of Fourier transform infrared spectroscopy to biomolecular profiling of cultured fibroblast cells from Gaucher disease patients: A preliminary investigation
| Autor: | Duygu Ozel Demiralp, Cemil Ozerk Demiralp, Serap Dökmeci, Aysel Yüce, Naşit İğci, Parisa Sharafi |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
Carbohydrates
Medicine (miscellaneous) 010501 environmental sciences 030226 pharmacology & pharmacy 01 natural sciences Protein Structure Secondary General Biochemistry Genetics and Molecular Biology 03 medical and health sciences 0302 clinical medicine Spectroscopy Fourier Transform Infrared Internal Medicine medicine Cluster Analysis Humans Pharmacology (medical) Fourier transform infrared spectroscopy Child Fibroblast Spectroscopy Protein secondary structure Cells Cultured Genetics (clinical) 0105 earth and related environmental sciences chemistry.chemical_classification Gaucher Disease Biomolecule Infant Proteins Enzyme replacement therapy Fibroblasts Lipids medicine.anatomical_structure Enzyme chemistry Biochemistry Child Preschool Reviews and References (medical) Glucocerebrosidase |
| Zdroj: | Advances in Clinical and Experimental Medicine. 26:1053-1061 |
| ISSN: | 1899-5276 |
| Popis: | BACKGROUND Gaucher disease (GD) is defined as an autosomal recessive disorder resulting from the deficiency of glucocerebrosidase (E.C. 3.2.1.45). Glucocerebrosidase is responsible for the degradation of glucosylceramide into ceramide and glucose. The deficiency of this enzyme results in the accumulation of undegraded glucosylceramide, almost exclusively in macrophages. With Fourier transform infrared (FTIR) spectroscopy, the complete molecular diversity of the samples can be studied comparatively and the amount of the particular materials can be determined. Also, the secondary structure ratios of proteins can be determined by analysing the amide peaks. OBJECTIVES The primary aim of this study is to introduce FTIR-ATR spectroscopy technique to GD research for the first time in the literature and to assess its potential as a new molecular method. MATERIAL AND METHODS Primary fibroblast cell cultures obtained from biopsy samples were used, since this material is widely used for the diagnosis of GD. Intact cells were placed onto a FTIR-ATR crystal and dried by purging nitrogen gas. Spectra were recorded in the mid-infrared region between 4500-850 cm-1 wavenumbers. Each peak in the spectra was assigned to as organic biomolecules according to their chemical bond information. A quantitative analysis was performed using peak areas and we also used a hierarchical cluster analysis as a multivariate spectral analysis. RESULTS We obtained FTIR spectra of fibroblast samples and assigned the biomolecule origins of the peaks. We observed individual heterogeneity in FTIR spectra of GD fibroblast samples, confirming the well-known phenotypic heterogeneity in GD at the molecular level. Significant alterations in protein, lipid and carbohydrate levels related to the enzyme replacement therapy were also observed, which is also supported by cluster analysis. CONCLUSIONS Our results showed that the application of FTIR spectroscopy to GD research deserves more attention and detailed studies with an increased sample size in order to evaluate its potential in the diagnosis and follow-up of GD patients. |
| Databáze: | OpenAIRE |
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