A comparative structural study in monolayers of GPI fragments and their binary mixtures.

Glycosylphosphatidylinositols (GPIs), natural complex glycolipids essential for a range of biological functions, are poorly understood with regard to their interactions and arrangements in cellular membranes. To evaluate the role of the head group in the structure formation in 2D model membranes (monolayers formed at the soft air/liquid interface), we employed the highly surface sensitive grazing incidence X-ray diffraction technique to investigate three GPI-fragments bearing the same hydrophobic part but different head groups. Condensed monolayers of simple GPI fragments are defined only by ordered alkyl chains. The monolayers of more complex fragments are additionally characterized by highly ordered head groups. Due to the strong H-bond network formed by the head groups, GPI-fragment both segregates and induces order into a model membrane phospholipid (POPC) that mimics the liquid-disordered phase of cell membranes. Here, we show that the strong van der Waals interactions between hydrophobic chains overcome the head group interactions and dominate the structure formation in mixtures of GPI-fragment with lipids that form liquid-condensed phases. This behaviour can be linked to the GPIs affinity for the lipid rafts.

Synthesis of glycosylphosphatidylinositol (GPI)-anchor glycolipids bearing unsaturated lipids.

2-Naphthyl-methyl ethers as permanent protecting groups are readily removed under acidic conditions and are key to the synthesis of complex glycosylphosphatidylinositol anchors containing unsaturated lipids. The total synthesis of the GPI pseudo-disaccharide core found on the surface of the Trypanosoma cruzi parasite serves to illustrate the power of the strategy.

Distinguishing N-acetylneuraminic acid linkage isomers on glycopeptides by ion mobility-mass spectrometry.

Differentiating the structure of isobaric glycopeptides represents a major challenge for mass spectrometry-based characterisation techniques. Here we show that the regiochemistry of the most common N-acetylneuraminic acid linkages of N-glycans can be identified in a site-specific manner from individual glycopeptides using ion mobility-mass spectrometry analysis of diagnostic fragment ions.

Quantitative mapping of glycoprotein micro-heterogeneity and macro-heterogeneity: an evaluation of mass spectrometry signal strengths using synthetic peptides and glycopeptides.

Mass spectrometry has made possible the field of proteomics and become an invaluable tool for identifying and quantifying post-translational modifications of proteins from complex mixtures. Because PTMs are recognized as key factors of biological activity, reliable PTM characterization is essential to understanding the relationship between protein isoform and activity. Protein glycosylations, in particular, can be especially difficult to characterize due to the range of different oligosaccharide entities that may be attached at any particular site. In this month' Special Feature Dr Daniel Kolarich of the Dept. of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces and his collaborators point out that quantitative label-free glycoproteomics can yield information about glycoprotein microand macroheterogeniety if the tools are sufficiently accurate. To understand and characterize the performance capability of MS tools they synthesized a panel of peptides and their glycopeptide derivatives as references and used these to investigate the qualitative and quantitative results from various ionization techniques and mass analyzers.