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 3 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 3 with lipids that form liquid-condensed phases. This behaviour can be linked to the GPIs affinity for the lipid rafts

Rescue of Glycosylphosphatidylinositol-Anchored Protein Biosynthesis Using Synthetic Glycosylphosphatidylinositol Oligosaccharides

The attachment of proteins to the cell membrane using a glycosylphosphatidylinositol (GPI) anchor is a ubiquitous process in eukaryotic cells. Deficiencies in the biosynthesis of GPIs and the concomitant production of GPI-anchored proteins lead to a series of rare and complicated disorders associated with inherited GPI deficiencies (IGDs) in humans. Currently, there is no treatment for patients suffering from IGDs. Here, we report the design, synthesis, and use of GPI fragments to rescue the biosynthesis of GPI-anchored proteins (GPI-APs) caused by mutation in genes involved in the assembly of GPI-glycolipids in cells. We demonstrated that the synthetic fragments GlcNAc-PI (1), Man-GlcN-PI (5), and GlcN-PI with two (3) and three lipid chains (4) rescue the deletion of the GPI biosynthesis in cells devoid of the PIGA, PIGL, and PIGW genes in vitro. The compounds allowed for concentration-dependent recovery of GPI biosynthesis and were highly active on the cytoplasmic face of the endoplasmic reticulum membrane. These synthetic molecules are leads for the development of treatments for IGDs and tools to study GPI-AP biosynthesis

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

Coherence length of an elongated condensate: a study by matter-wave interferometry

We measure the spatial correlation function of Bose-Einstein condensates in the cross-over region between phase-coherent and strongly phase-fluctuating condensates. We observe the continuous path from a gaussian-like shape to an exponential-like shape characteristic of one-dimensional phase-fluctuations. The width of the spatial correlation function as a function of the temperature shows that the condensate coherence length undergoes no sharp transition between these two regimes.Comment: 8 pages, 6 figure, submitted to EPJ

Quantum Gates and Memory using Microwave Dressed States

Trapped atomic ions have been successfully used for demonstrating basic elements of universal quantum information processing (QIP). Nevertheless, scaling up of these methods and techniques to achieve large scale universal QIP, or more specialized quantum simulations remains challenging. The use of easily controllable and stable microwave sources instead of complex laser systems on the other hand promises to remove obstacles to scalability. Important remaining drawbacks in this approach are the use of magnetic field sensitive states, which shorten coherence times considerably, and the requirement to create large stable magnetic field gradients. Here, we present theoretically a novel approach based on dressing magnetic field sensitive states with microwave fields which addresses both issues and permits fast quantum logic. We experimentally demonstrate basic building blocks of this scheme to show that these dressed states are long-lived and coherence times are increased by more than two orders of magnitude compared to bare magnetic field sensitive states. This changes decisively the prospect of microwave-driven ion trap QIP and offers a new route to extend coherence times for all systems that suffer from magnetic noise such as neutral atoms, NV-centres, quantum dots, or circuit-QED systems.Comment: 9 pages, 4 figure

COPII coat composition is actively regulated by luminal cargo maturation

Background Export from the ER is an essential process driven by the COPII coat, which forms vesicles at ER exit sites (ERESs) to transport mature secretory proteins to the Golgi. Although the basic mechanism of COPII assembly is known, how COPII machinery is regulated to meet varying cellular secretory demands is unclear. Results Here, we report a specialized COPII system that is actively recruited by luminal cargo maturation. Glycosylphosphatidylinositol-anchored proteins (GPI-APs) are luminal secretory proteins anchored to the membrane by the glycolipid GPI. After protein attachment in the ER lumen, lipid and glycan parts of the GPI anchor are remodeled. In yeast, GPI-lipid remodeling concentrates GPI-APs into specific ERESs. We found that GPI-glycan remodeling induces subsequent recruitment of the specialized ER export machinery that enables vesicle formation from these specific ERESs. First, the transmembrane cargo receptor p24 complex binds GPI-APs as a lectin by recognizing the remodeled GPI-glycan. Binding of remodeled cargo induces the p24 complex to recruit the COPII subtype Lst1p, specifically required for GPI-AP ER export. Conclusions Our results show that COPII coat recruitment by cargo receptors is not constitutive but instead is actively regulated by binding of mature ligands. Therefore, we reveal a novel functional link between luminal cargo maturation and COPII vesicle budding, providing a mechanism to adjust specialized COPII vesicle production to the amount and quality of their luminal cargos that are ready for ER exit. This helps to understand how the ER export machinery adapts to different needs for luminal cargo secretion.This work was supported by grants from the Spanish Ministry of Science and Innovation BFU2008-04119 (to M.M.), BFU2011-24513 (to M.M.), BFU2009-07290 (to V.G.), and BFU2010-21339 (to R.E.W.); Junta de Andalucia P09-CVI-4503 (to M.M.) and P11-CTS-7962 (to R.E.W.); Swiss National Science Foundation and the NCCR Chemical Biology (to H.R.); the Max-Planck Society (to P.H.S.); the RIKEN-Max-Planck Joint Center for Chemical Biology (to D.V.S.); by JSPS KAKENHI grant number 21580094 (to K.F.); University of Seville fellowships (to A.A.-R., J.M.-L., and A.M.P.-L.); and Ramon y Cajal program (to V.G.). We are grateful to Servicios de Biología y Microscopía (CITIUS, Universidad de Sevilla)