Separation of hyaluronan oligosaccharides by the use of anion-exchange HPLC

Holmbeck, S. M. A.; Lerner, Laura

doi:10.1016/0008-6215(93)84218-u

Cited by 20 publications

(8 citation statements)

References 21 publications

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“…These included 2,5-dihydroxybenzoic acid (DHBA), which has been used recently for the analysis of HA oligomers [30,31], but this matrix did not generate the strong signals previously reported by Mahoney et al [32]. In the positiveion mass spectra, the major peaks were generally a mixture of metal ion adducts, such as [M +Na] + and [M+K] + , whereas [M−H] − was the predominant species in the negative-ion mode (data not shown) as described previously [33]. Consequently, a new strategy was designed to control the formation of counter-ion species of uronate residues in the MALDI-TOF mass spectra of HA oligosaccharides.…”

Section: Comparison Of Ha Oligosaccharides Salts and Methylester For supporting

confidence: 58%

Matrix assisted laser desorption ionization-time of flight mass spectrometry analysis of hyaluronan oligosaccharides

Sakai

Hirano

Toyoda

et al. 2007

Analytica Chimica Acta

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A new method is presented for the identification of oligosaccharides obtained by enzymatic digestion of hyaluronan (HA) with bacterial hyaluronidase (E.C. 4.2.2.1, from Streptomyces hyalurolyticus) using matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOFMS). Mixtures containing HA oligosaccharides of tetrasaccharide (4-mer)-34-mer were analyzed using this method. The carboxyl groups of the glucuronate residues in the prepared HA oligomers, were modified as the acidic form (-COOH), sodium salts (-COONa), organic ammonium salts, or methylesters before MALDI-TOFMS measurement. Among these samples, the methylester form of glucuronate residues in HA oligosaccharides, prepared by methylation using trimethylsilyl diazomethane, afforded high sensitivity for spectra. This simple modification method for carboxyl group methylation of acidic polysaccharides [Hirano et al., Carbohydr. Res., 340, (2005) 2297-2304] provides samples suitable for MALDI-TOF mass spectrometric analysis throughout a significantly enhanced range of masses.

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Section: Comparison Of Ha Oligosaccharides Salts and Methylester For supporting

confidence: 58%

Matrix assisted laser desorption ionization-time of flight mass spectrometry analysis of hyaluronan oligosaccharides

Sakai

Hirano

Toyoda

et al. 2007

Analytica Chimica Acta

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“…Sample Preparation. The hyaluronan octasaccharide containing four repeating disaccharide units (HA4), with /3-dglucuronate (GlcA) on the nonreducing end and 2-acetamido-2-deoxy-D-glucose GV-acetylglucosamine, GlcNAc) on the reducing end, was prepared by enzymatic degradation of high molecular weight hyaluronan, as described in previous work (Holmbeck & Lemer, 1993).…”

Section: Methodsmentioning

confidence: 99%

The solution conformation of hyaluronan: A combined NMR and molecular dynamics study

Holmbeck¹,

Petillo²,

Lerner³

1994

Biochemistry

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Hyaluronan (HA) is a negatively charged glycosaminoglycan that exhibits a wide variety of biological effects mediated by binding to cell-surface and extracellular matrix proteins (hyaladherins). Short HA oligosaccharides have been shown to retain the specific interactions and biological effects of high molecular weight HA. Although it has a simple disaccharide repeating unit, the aqueous solution conformation of HA has been very difficult to determine because of strong coupling and overlapping resonances. In this study, we propose aqueous solution conformations for an octasaccharide of HA, derived from proton-proton NOE data and restrained molecular dynamics. To overcome spectral overlap and strong coupling, alternate methods for extracting distance restraints were employed. Restrained molecular dynamics calculations yielded one set of interglycosidic angle values for the beta (1,3) linkage (phi 13 = 46 degrees, psi 13 = 24 degrees). In contrast, two sets of values for the beta (1,4) linkage were consistent with the NOE restraints (phi 14 = 24 degrees, psi 14 = -53 degrees or phi 14 = 48 degrees, psi 14 = 8 degrees). The potential difference in flexibility for the two linkages is consistent with unrestrained as well as the restrained molecular dynamics trajectories described here. The conformational parameters obtained from restrained molecular dynamics are used to predict helical parameters of high molecular weight HA and will provide a basis for studies of HA binding to proteins.

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“…After heat inactivation of the enzyme, the oligosaccharide solution was sterile-filtered twice using a 0.2-m Nalgene disposable tissue culture filter. Parallel experiments determined the sizes of the oligosaccharides by separating the digested mixture using a DEAE ion exchange column and a quantitative carbazole-based assay of HA with a reducing power assay (data not shown; [43,44]). To ensure that a substantial amount of HA oligosaccharides would be present at the time of I␣I and HA binding (ϳ6 h post-hCG), the half-life of the oligosaccharides was quantitated using the Bitter and Muir [45] carbazole assay.…”

Section: Ha Oligosaccharidesmentioning

confidence: 99%

“…Previous studies using the methods of Hascall and Heinegard [42] and Holmbeck and Lerner [43], with minor modifications, have shown that a solution of HA oligosaccharides (5-100 disaccharides) can be produced by partial degradation of rooster comb HA with testicular hyaluronidase. After heat inactivation of the enzyme, the oligosaccharide solution was sterile-filtered twice using a 0.2-m Nalgene disposable tissue culture filter.…”

Section: Ha Oligosaccharidesmentioning

confidence: 99%

Inter-α-Inhibitor Binding to Hyaluronan in the Cumulus Extracellular Matrix Is Required for Optimal Ovulation and Development of Mouse Oocytes1

Hess

Chen

Larsen

1999

Biology of Reproduction

114

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This report characterizes the effects of excess hyaluronan (HA) upon the expansion of the cumulus oocyte complex (COC) within intact follicles and upon ovulation and oocyte viability in mice. Covalent linkage between heavy chains of the inter-alpha-inhibitor (IalphaI) family of serum glycoproteins and HA is necessary for optimal cumulus extracellular matrix (cECM) stabilization and cumulus expansion. Intravenous administration of HA oligosaccharides inhibited the binding of IalphaI to endogenous HA, disrupting the process of expansion and resulting in a reduction in the size of the cumulus mass. Western blot and immunocytochemical analyses of COCs from HA-treated animals demonstrated a reduction of IalphaI heavy chains within the cECM. Additionally, HA-treated immature animals ovulated 56.3% fewer COCs compared to control animals. The developmental potential of COCs in HA-treated animals was also tested. Extended periods of oviductal storage of COCs ovulated by HA-injected adult mice resulted in a reduction of normal embryos and a significant increase in the proportion of fragmented oocytes/embryos. These observations support the view that covalent binding of IalphaI heavy chains to HA is required for optimal cumulus expansion, extrusion of the COCs from the follicle at ovulation, and maintenance of oocyte viability within the oviduct.

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Separation of hyaluronan oligosaccharides by the use of anion-exchange HPLC

Cited by 20 publications

References 21 publications

Matrix assisted laser desorption ionization-time of flight mass spectrometry analysis of hyaluronan oligosaccharides

Matrix assisted laser desorption ionization-time of flight mass spectrometry analysis of hyaluronan oligosaccharides

The solution conformation of hyaluronan: A combined NMR and molecular dynamics study

Inter-α-Inhibitor Binding to Hyaluronan in the Cumulus Extracellular Matrix Is Required for Optimal Ovulation and Development of Mouse Oocytes1

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