High performance mass spectrometry for advanced interactomics studies

Robu Adrian C. and Popescu Laurentiu and Seidler Daniela G. and Zamfir Alina D.: High performance mass spectrometry for advanced interactomics studies.

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Fibroblast growth factor-2 (FGF-2) is a glycosaminoglycan (GAG) binding protein, involved in different biological processes, such as angiogenesis, bone signaling, embryonic development, morphogenesis or cartilage metabolism. GAGs, one of its binding partners, are long-unbranched polysaccharides exhibiting a repeating disaccharide unit. Moreover, preceding studies have shown that GAGs play an important role in tissue development, cellular behavior or extracellular matrix (ECM) organization. The FGF-GAG noncovalent interactions are of high importance in the biological and biomedical fields of research, as a result of their influence in the tissue regeneration and cell proliferation processes. Here, we have employed one of the most advanced mass spectrometric (MS) techniques consisting of fully automated chip-nanoelectrospray (nanoESI), coupled to a quadrupole time-of-flight (QTOF) MS for studying the FGF-GAG noncovalent complexes. The experiments were conducted in 10 mM ammonium acetate/formic acid, pH 6.8, by incubating FGF-2 and CS disaccharides dissolved in buffer; aliquots were collected after 5, 10, 30, 60 and 90 minutes and further submitted to chip-based MS analysis. For the first time, a CS disaccharide was involved in a binding assay with FGF-2. The detected complexes in the screening experiments were further characterized by top-down fragmentation in tandem MS (MS/MS) using collision induced-dissociation (CID) at low ion acceleration energies. CID MS/MS provided data showing for the first time that the binding process occurs via SO3 located at C4 in the GalNAc moiety.

Item Type: Conference or Workshop Item
Journal or Publication Title: Proceedings of the International Symposium on Analytical and Environmental Problems
Date: 2017
Volume: 23
ISBN: 978-963-306-563-1
Page Range: p. 305
Event Title: International Symposium on Analytical and Environmental Problems (23.) (2017) (Szeged)
Related URLs: http://acta.bibl.u-szeged.hu/55894/
Uncontrolled Keywords: Elektrokémia - előadáskivonat
Date Deposited: 2019. Jan. 02. 11:25
Last Modified: 2022. Aug. 08. 15:49
URI: http://acta.bibl.u-szeged.hu/id/eprint/56211

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