Glycopeptides characterization

Delfour, A., Jolles, J., Alais, C., and Jolles, P. (1965). Caseino-glycopeptides Characterization of a methionine residue and of the N-terminal sequence. Biochem. Biophys. Res. Commun. 19,452-455. 

Kits for analyzing glycoproteins are now available, utilizing recombinant endo-enzymes together with electrophoresis and fluorescent labeling with ANTS (aminonaphthalene-l,3,6-trisulfonic acid) PMP (1-phenyl-3-methyl-5-pyrazolone), or other, similar type of reagent can be used. MS plays a crucial role in glycopeptide characterization where techniques such as electrospray LC-MS and MALDI-TOF MS are utilized. 

Rohrer, J. S., Cooper, G. A., and Townsend, R. R., Identification, quantification, and characterization of glycopeptides in reversed-phase HPLC separations of glycoprotein proteolytic digests, Anal. Biodiem., 212, 7, 1993. 

The N-linked pentasaccharide core Man3(GlcNAc)2 glycopeptide bearing the extracellular MMP inducer sequence 517 (emmprin 34—58) has been successfully linked to G(l) PAMAM aminodendrimer by thioester activation (Fig. 63). The resulting octameric 30 kDa construct was obtained in low yield, but was purified by preparative electrophoresis and fully characterized by MALDI-TOF mass spectrometry. The multivalent architecture was built to evaluate the requirement of emmprin multimerization for inducing MMP expression.384... 

For novel oligosaccharides for which H-n.m.r. data have not previously been documented in the literature, it is necessary to supplement the H-n.m.r. data with composition, sequence, and linkage information provided by chromatographic and m.s. analyses. An illustrative example is the characterization of the following oligosaccharide of the glycopeptides of fetal gastrointestinal mucins of meconium (74) ... 

The development glycopeptide libraries obtained by the split-mix method is severely hampered by the lack of concurrent development of a general, facile separation and characterization technology. Some headway has been made with chemical coding of the libraries, but very few direct methods of analysis exist. One promising method that could be applied to the direct characterization of both types of libraries is mass spectrometry. More specifically, post-source-decay matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (PSD-MALDI-TOF-MS) and CID-FAB/MS/MS have been used to characterize glycopeptides.53-55... 

Arthur, M. Molinas, C. Depardieu, E Courvalin, P Characterization of Tnl546, a Tn3-related transposon conferring glycopeptide resistance by synthesis of depsipeptide peptidoglycan precursors in Enterococcus faecium BM4147. J. Bacteriol., 175, 117-127 (1993)... 

Fig. 25 Characterization and luciferase expression of PGP DNA condensates in vivo. These results show that luciferase expression is dependent on galactose incorporation but independent of amount of melittin. (a) Represents the input mol ratio of Cys-terminated melittin, PEG-peptide, and glycopeptide. (b) Represents the measured mol ratio of Cys-terminated melittin, PEG-peptide, and glycopeptide for each purified PGP. (c) Values are the calculated MW based on polylysine standards, (d) Values are the calculated MW based on PEG standards, (e) The mean particle size determined at a stoichiometry of 0.3 nmol of PGP per mg of DNA. The value represents the mean diameter (nm) based on unimodal analysis, (f) The zeta potential of PGP DNA condensates at a stoichiometry of 0.3 nmol of PGP per mg of DNA. (g) The metabolic half-life of PGP 125I-DNA in triplicate mice. The results are derived from Fig. 6. (h) The PC/NPC ratio of DNA-targeted liver, (i) Represents a control PGP 3 in which galactose has been removed. Figure adapted with permission from [182], 2007 American Chemical Society...

Subsequent experimental work in this laboratory was aimed at the systematic development of an efficient method for isolating the proteinaceous surfactants, which help stabilize natural microbubbles, from both commercial agarose powder and from forest soil samples collected locally. Successful isolation of this glycopeptide fraction was eventually achieved (ref. 322), and the results obtained from an extended program of chemical analysis, to further characterize and compare chemically these proteinaceous surfactants from both natural substances, are described below. 

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