... -treated LiP) was inactive. MS analyses revealed that VA-LiP includes one VA molecule covalently bound to the side chain of Tyr 181 , whereas H 2 O 2 -LiP contains a hydroxylated Tyr 181 . No adduct is formed in the Y171N variant. Molecular docking showed that VA binding is favoured by sandwich π stacking...

... of expression of OAT2 is the sinusoidal membrane domain of hepatocytes. The particular physiological function of OAT2 in liver has been unresolved so far. In the present paper, we have used the strategy of LC (liquid chromatography)–MS difference shading to search for specific and cross-species substrates...

... in solution which are often difficult to quantify. In the present manuscript we show that the combination of MS under nondenaturing conditions and AUC-SV (analytical ultracentrifugation sedimentation velocity) unambiguously clarifies a distribution of states and hydrodynamic shapes of assembled oligomers...

... and then analysed by LC–ESI–MS/MS (liquid chromatography–electrospray ionization–tandem MS). A total of 93 proteins were identified. Among the proteins identified, we concentrated on the E3 ubiquitin-protein ligase Nedd4 (neural-precursor-cell-expressed, developmentally down-regulated gene 4), and performed...

... confirmed that the C-terminal tail of MUC17 co-precipitates with the scaffold protein PDZK1 as identified by MS. This was mediated through the C-terminal PDZ-interaction site in MUC17, which was capable of binding to three of the four PDZ domains in PDZK1. Immunostaining of wild-type or Pdzk1 −/− mouse...

... posttranslational modifications by MS. We have found that Max is acetylated at several lysine residues (Lys-57, Lys-144 and Lys-145) in mammalian cells. Max acetylation is stimulated by inhibitors of histone deacetylases and by overexpression of the p300 co-activator/HAT (histone acetyltransferase). The p300 HAT...

... sulphation levels, which allows cells to tune their affinities to various extracellular proteins, including growth factors. With stable isotope labelling and HPLC-coupled MS, modification degrees at various O-sulphation sites could be determined. A bovine kidney HS sample was first saturated in vitro with 34...

...). Fatty acid-free or -bound HSA was modified with lysine-specific cross-linkers and digested with trypsin. Cross-linked peptides were analysed by nano-electrospray ionization MS to localize the sites of cross-linking. Our data indicated that a local conformational change involving movement of the side...

... are expressed in human epithelial, glioma and promyelocytic cell lines; (ii) NATH and hARD1 form a stable complex, as investigated by reciprocal immunoprecipitations followed by MS analysis; (iii) NATH–hARD1 complex expresses N-terminal acetylation activity; (iv) NATH and hARD1 interact with ribosomal subunits...

... medium, in cytosol of Kato III cells and in cytosol of erythrocytes. GSTP1 subunits from Kato III cells and erythrocytes were heterogeneous when analysed by MALDI–TOF (matrix-assisted laser-desorption ionization–time-of-flight) MS, suggesting that there were post-translational modifications to GSTP1. One...

... depends on the carbohydrate content and glycoform distribution, we performed an MS analysis of the protein sugar moieties. Glycosidic structures of the high mannose type were found, with mannoses ranging from 8 to 25 residues. 1 To whom correspondence should be addressed (email silviamaria.doglia...

..., dried and analysed by MALDI–TOF-MS. Mild acid hydrolysis was performed using 2 M HCl at 80 °C for 30 min. NMR spectra were acquired in 2 H 2 O and recorded at 300 K. 1 H-NMR spectra were obtained at 400 MHz using a Varian Unity 400 instrument. 13 C and two-dimensional NMR spectra were obtained...

... treatment the released oligosaccharides were structurally characterized by nano-ESI Q-TOF MS/MS (electrospray ionization quadrupole time-of-flight tandem MS) without prior fractionation or derivatization. More than 100 different oligosaccharides, with up to ten monosaccharide residues, were identified using...

... aprotinin, 1 μg/ml antipain, chymostatin and leupeptin). chaperone complex I assembly MS NADH dehydrogenase prohibitin NADH:ubiquinone oxidoreductase (complex I, EC 1.6.5.3) catalyses the oxidation of NADH, with the concomitant reduction of ubiquinone and ejection of protons out...

... was studied further by mutational analyses and by using reversed-phase HPLC and MS. The reversed-phase HPLC elution pattern of the three isoforms revealed that Ser-650 was phosphorylated constitutively. After de novo synthesis, only Ser-650 was phosphorylated in the smallest isoform of 110 kDa and both Ser...

... identification of proteins at the opposite membrane surfaces by proteolysis and MS. Extracellular-exposed caveolae-specific proteins CD36 and copper-containing amine oxidase were concealed inside the vesicles and resisted trypsin treatment. The cytosol-orientated caveolins were efficiently digested by trypsin...

... a highly developed MS strategy to identify the cdk5 phosphorylation sites and determine the major and minor sites. Several phosphopeptides were identified from a tryptic digest of 32 P-labelled, cdk5-phosphorylated DCX using a combination of off-line HPLC and matrix-assisted laser-desorption ionization-MS...

... identified by MALDI-TOF (matrix-assisted laser-desorption ionization–time-of-flight) MS as calreticulin, nucleophosmin, phosphoglycerate kinase and pyruvate kinase. How the up-regulation of these proteins might help to explain the modification of Bcl2 activity is discussed. 1 These authors contributed...

... ). 29 10 2002 11 2 2003 27 2 2003 27 2 2003 The Biochemical Society, London ©2003 2003 glycosylphosphatidylinositol anchor MS membrane soluble ecto-5′-nucleotidase Abbreviations used: GPI, glycosylphospatidylinositol; IEF, isoelectric focusing; MALDI, matrix...

... are mediated by intracellular metabolites. 1 To whom correspondence should be addressed (e-mail [email protected] ). 20 12 2002 28 1 2003 11 2 2003 11 2 2003 The Biochemical Society, London ©2003 2003 fibroblast flavonoid glucuronide MS O -methylated...

... mucins of the small intestine in CF mice revealed a large increase in fucose, whereas those of wild-type mice contained only small amounts of fucose. This increased fucosylation was analysed by releasing the O-linked oligosaccharides followed by GC-MS. NMR spectroscopy revealed that the increased...

...Åsa C. HAGLUND; Bo EK; Pia EK A fraction of α 2 -Heremans–Schmid (α 2 -HS) glycoprotein (human fetuin) isolated from plasma was phosphorylated at serine-120 and serine-312 as shown by MS and peptide fragment sequencing after tryptic digestion. Serine-312-containing peptides were phosphorylated...

...William J. GRIFFITHS; Andreas P. JONSSON; Suya LIU; Dilip K. RAI; Yuqin WANG Over the last 20 years, biological MS has changed out of all recognition. This is primarily due to the development in the 1980s of ‘soft ionization’ methods that permit the ionization and vaporization of large, polar...