High-resolution mass spectrometry reveals a complex ganglioside pattern and novel polysialylated structures associated with the human motor cortex

Abstract

We have developed here a superior methodology based on high-resolution mass spectrometry for screening and fragmentation analysis of gangliosides extracted and purified from the human motor cortex . The experiments, conducted on a nanoelectrospray Orbitrap mass spectroscope in the negative ion mode, allowed the discrimination in the native mixture extracted from human motor cortex of no less than 83 different gangliosides, which represents the highest number of structures identified so far in this brain region. The spectral data, acquired in high-resolution mass spectrometry mode with a remarkable sensitivity and an average mass accuracy of 4.48 ppm, also show that the gangliosidome of motor cortex is generally characterized by species exhibiting a much higher degree of sialylation than previously known. Motor cortex was found dominated by complex structures with a sialylation degree ≥3, exhibiting long saccharide chains, in the G1 class. Fucogangliosides and species with the glycan chain elongated by either O-acetylation and/or acetate anion attachments were also detected; the later modification was for the first time discovered in this brain region. Of major significance is the identification of hepta and octasialylated species of GS1 and GO1 type, which are among the structures with the longest oligosaccharide chain discovered so far in the human brain. In the last stage of research, tandem mass spectrometry performed by higher energy collision dissociation provided structural data documenting the occurrence of GT1b (d18:1/20:0) isomer in the human motor cortex.

Advances in Mass Spectrometry of Gangliosides Expressed in Brain Cancers

“…The development, a few years later, of nanoelectrospray (nanoESI) and matrix-assisted laser desorption/ionization (MALDI) methods represented the factual breakthrough of mass spectrometry in human brain ganglioside analysis [32][33][34]. With the succeeding technical innovations, such as microfluidics-MS [34][35][36][37][38], the high resolution analyzers, such as Fourier transform ion cyclotron resonance (FTICR) [39,40], Orbitrap [41,42], ion mobility separation (IMS) MS [43,44] as well as desorption electrospray (DESI) and MALDI MS imaging (MSI) [45][46][47] and the efficient ion fragmentation methods [48], the achievements of mass spectrometry in profiling, detailed structural analysis and discovery of normal and pathological brain-associated gangliosides increased spectacularly. In this context, the present review discusses the trends in the important field of brain cancer ganglioside analysis using modern MS approaches, highlighting the invaluable contributions of this method in establishing well-defined sets of biomarkers and elucidating some of the ganglioside-dependent mechanisms of tumor proliferation.…”