Application of electrospray mass spectrometry to the structural determination of glycosylphosphatidylinositol membrane anchors. 2010

Isabelle R E Nett, and Angela Mehlert, and Douglas Lamont, and Michael A J Ferguson
Division of Biological Chemistry and Drug Discovery, College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK.

The addition of glycosylphosphatidylinositol (GPI) anchors to proteins is an important posttranslational modification in eukaryotic cells. The complete structural elucidation of GPI anchors is a complex process that requires relatively large amounts of starting material. In this paper, we assess the degree of structural information that can be obtained by applying electrospray mass spectrometry and tandem mass spectrometry to permethylated GPI glycans prepared from a well-characterized GPI-anchored glycoprotein, the variant surface glycoprotein from Trypanosoma brucei. All GPI glycans contain a non-N-acetylated glucosamine residue, and permethylation leads to the formation of a fixed positive charge on the glycans, in the form of a quaternary amine. The permethylated glycans were detected as [M +- Na](2+-) ions, and tandem mass spectrometry of these ions produced substantial, albeit incomplete, structural information on the branching patterns and linkage types for various GPI glycoforms of the variant surface glycoprotein.

UI MeSH Term Description Entries
D011134 Polysaccharides Long chain polymeric CARBOHYDRATES composed of MONOSACCHARIDES linked by glycosidic bonds. Glycan,Glycans,Polysaccharide
D014346 Trypanosoma brucei brucei A hemoflagellate subspecies of parasitic protozoa that causes nagana in domestic and game animals in Africa. It apparently does not infect humans. It is transmitted by bites of tsetse flies (Glossina). Trypanosoma brucei,Trypanosoma brucei bruceus,Trypanosoma bruceus,brucei brucei, Trypanosoma,brucei, Trypanosoma brucei,bruceus, Trypanosoma,bruceus, Trypanosoma brucei
D014643 Variant Surface Glycoproteins, Trypanosoma Glycoproteins attached to the surface coat of the trypanosome. Many of these glycoproteins show amino acid sequence diversity expressed as antigenic variations. This continuous development of antigenically distinct variants in the course of infection ensures that some trypanosomes always survive the development of immune response to propagate the infection. Surface Variant Glycoproteins, Trypanosoma,Trypanosoma Variant Surface Coat Glycoproteins,SSP-4,VSG 117,VSG 118,VSG 221
D015394 Molecular Structure The location of the atoms, groups or ions relative to one another in a molecule, as well as the number, type and location of covalent bonds. Structure, Molecular,Molecular Structures,Structures, Molecular
D017261 Glycosylphosphatidylinositols Compounds containing carbohydrate or glycosyl groups linked to phosphatidylinositols. They anchor GPI-LINKED PROTEINS or polysaccharides to cell membranes. GPI Membrane Anchor,GPI Membrane Anchors,Glycosyl-Phosphatidylinositol Membrane Protein Anchor,Glycosylated Phosphatidylinositol,Glycosylphosphatidylinositol Anchor,Glycosylphosphatidylinositol Anchors,Phosphatidylinositol Glycan,Gly-PtdIns,Glycoinositol Phospholipid Membrane Anchor,Glycosyl-Phosphatidylinositol,Glycosyl-Phosphatidylinositol Membrane Protein Anchors,Glycosylated Phosphatidylinositols,Glycosylphosphatidylinositol,Glycosylphosphatidylinositol Linkage,PI-Glycan,Anchor, GPI Membrane,Anchor, Glycosylphosphatidylinositol,Anchors, GPI Membrane,Anchors, Glycosylphosphatidylinositol,Glycan, Phosphatidylinositol,Glycosyl Phosphatidylinositol,Glycosyl Phosphatidylinositol Membrane Protein Anchor,Glycosyl Phosphatidylinositol Membrane Protein Anchors,Linkage, Glycosylphosphatidylinositol,Membrane Anchor, GPI,Membrane Anchors, GPI,PI Glycan,Phosphatidylinositol, Glycosylated,Phosphatidylinositols, Glycosylated
D053719 Tandem Mass Spectrometry A mass spectrometry technique using two (MS/MS) or more mass analyzers. With two in tandem, the precursor ions are mass-selected by a first mass analyzer, and focused into a collision region where they are then fragmented into product ions which are then characterized by a second mass analyzer. A variety of techniques are used to separate the compounds, ionize them, and introduce them to the first mass analyzer. For example, for in GC-MS/MS, GAS CHROMATOGRAPHY-MASS SPECTROMETRY is involved in separating relatively small compounds by GAS CHROMATOGRAPHY prior to injecting them into an ionization chamber for the mass selection. Mass Spectrometry-Mass Spectrometry,Mass Spectrometry Mass Spectrometry,Mass Spectrometry, Tandem
D021241 Spectrometry, Mass, Electrospray Ionization A mass spectrometry technique used for analysis of nonvolatile compounds such as proteins and macromolecules. The technique involves preparing electrically charged droplets from analyte molecules dissolved in solvent. The electrically charged droplets enter a vacuum chamber where the solvent is evaporated. Evaporation of solvent reduces the droplet size, thereby increasing the coulombic repulsion within the droplet. As the charged droplets get smaller, the excess charge within them causes them to disintegrate and release analyte molecules. The volatilized analyte molecules are then analyzed by mass spectrometry. ESI Mass Spectrometry,Electrospray Ionization Mass Spectrometry,Mass Spectrometry, ESI,Spectrometry, ESI Mass

Related Publications

Isabelle R E Nett, and Angela Mehlert, and Douglas Lamont, and Michael A J Ferguson
June 1994, Glycoconjugate journal,
Isabelle R E Nett, and Angela Mehlert, and Douglas Lamont, and Michael A J Ferguson
January 1994, Methods in enzymology,
Isabelle R E Nett, and Angela Mehlert, and Douglas Lamont, and Michael A J Ferguson
September 1995, The Biochemical journal,
Isabelle R E Nett, and Angela Mehlert, and Douglas Lamont, and Michael A J Ferguson
May 2000, Journal of the American Society for Mass Spectrometry,
Isabelle R E Nett, and Angela Mehlert, and Douglas Lamont, and Michael A J Ferguson
April 1999, Seminars in perinatology,
Isabelle R E Nett, and Angela Mehlert, and Douglas Lamont, and Michael A J Ferguson
December 1993, The Journal of biological chemistry,
Isabelle R E Nett, and Angela Mehlert, and Douglas Lamont, and Michael A J Ferguson
March 2003, Glycobiology,
Isabelle R E Nett, and Angela Mehlert, and Douglas Lamont, and Michael A J Ferguson
December 1995, Journal of the American Society for Mass Spectrometry,
Isabelle R E Nett, and Angela Mehlert, and Douglas Lamont, and Michael A J Ferguson
April 2005, Journal of mass spectrometry : JMS,
Isabelle R E Nett, and Angela Mehlert, and Douglas Lamont, and Michael A J Ferguson
August 1999, Journal of chromatography. A,
Copied contents to your clipboard!