The glycosyl-phosphatidylinositol anchor of membrane proteins.

The only identified component of the scrapie prion is PrPSc, a glycosylinositol phospholipid (GPI)-linked protein that is derived from the cellular isoform (PrPC) by an as yet unknown posttranslational event. Analysis of the PrPSc GPI has revealed six different glycoforms, three of which are unprecedented. Two of the glycoforms contain N-acetylneuraminic acid, which has not been previously reported as a component of any GPI. The largest form of the GPI is proposed to have a glycan core consisting of Man alpha-Man alpha-Man-(NeuAc-Gal-GalNAc-)Man-GlcN-Ino. Identical PrPSc GPI structures were found for two distinct isolates or "strains" of prions which specify different incubation times, neuropathology, and PrPSc distribution in brains of Syrian hamsters. Limited analysis of the PrPC GPI reveals that it also has sialylated glycoforms, arguing that the presence of this monosaccharide does not distinguish PrPC from PrPSc.

Glycosylinositol phospholipid anchors of the scrapie and cellular prion proteins contain sialic acid.

Mycobacterium tuberculosis is the etiologic agent of tuberculosis and can be accurately detected by laboratories using commercial genetic tests. Nontuberculosis mycobacteria (NTM) causing other mycobacterioses can be difficult to identify. The identification processes are confounded by an increasing diversity of newly characterized NTM species. The ubiquitous nature of NTM, combined with their potential to be opportunistic pathogens in immunocompromised as well as nonimmunodeficient patients, further complicates the problem of their identification. Since clinical case management varies depending on the etiologic agent, laboratories must identify the species in a timely manner. However, only a few identification methods can detect the species diversity within the Mycobacterium genus. Over the last decade, high-performance liquid chromatography analysis of the mycolic acids has become an accepted method for identification of mycobacteria. In this review, we assess its development and usefulness as an identification technique for Mycobacterium species.

https://cmr.asm.org/content/cmr/14/4/704.full.pdf

Mycolic Acid Analysis by High-Performance Liquid Chromatography for Identification of Mycobacterium Species

Fast atom bombardment mass spectrometry for structural elucidation of glycoconjugates

A 125-kDa glycoprotein exposed on the surface of Saccharomyces cerevisiae cells belongs to a class of eucaryotic membrane proteins anchored to the lipid bilayer by covalent linkage to an inositol-containing glycophospholipid. We have cloned the gene (GAS1) encoding the 125-kDa protein (Gas1p) and found that the function of Gas1p is not essential for cell viability. The nucleotide sequence of GAS1 predicts a 60-kDa polypeptide with a cleavable N-terminal signal sequence, potential sites for N- and O-linked glycosylation, and a C-terminal hydrophobic domain. Determination of the anchor attachment site revealed that the C-terminal hydrophobic domain of Gas1p is removed during anchor addition. However, this domain is essential for addition of the glycophospholipid anchor, since a truncated form of the protein failed to become attached to the membrane. Anchor addition was also abolished by a point mutation affecting the hydrophobic character of the C-terminal sequence. We conclude that glycophospholipid anchoring of Gas1p depends on the integrity of the C-terminal hydrophobic domain that is removed during anchor attachment.

Determinants for glycophospholipid anchoring of the Saccharomyces cerevisiae GAS1 protein to the plasma membrane.

Mass spectrometric localization of carbon-carbon double bonds: A critical review of recent methods

Current methods are reviewed for determining the position of double bonds in fatty acids, and other unsaturated organic compounds, using mass spectrometry. ‘On-site’ and ‘remote-site’ derivatization methods are described, and their advantages and disadvantages for mass spectrometric analysis discussed. Chemical transformation of double bonds by methoxylation, silyloxylation or deuteration, together with electron impact (EI), chemical ionization (CI) or collisionally induced decomposition (CID) techniques in combination with fast atom bombardment (FAB) or CI, are found to be most suitable for polyunsaturated fatty acids (PUFA): for the analysis of less unsaturated compounds on a submicrogram scale those methods are most promising which either do not involve derivatization of the double bonds, or give derivatives in quantitative yields. Effects of mass spectrometer geometry and operating conditions are also considered.

Mass Spectrometric Localization of Carbon-Carbon Double Bonds: A Critical Review of Recent Methods

MS and NMR analysis of the cross-reacting determinant glycan from Trypanosoma brucei brucei MITat 1.6 variant specific glycoprotein.

Tyrosine 734 of NCAM180 interferes with FGF receptor-dependent signaling implicated in neurite growth

Topics in lipid research : from structural elucidation to biological function

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