Novel methods for the preparation and characterization of hyaluronan oligosaccharides of defined length.
Identifieur interne : 003431 ( Main/Exploration ); précédent : 003430; suivant : 003432Novel methods for the preparation and characterization of hyaluronan oligosaccharides of defined length.
Auteurs : D J Mahoney [Royaume-Uni] ; R T Aplin ; A. Calabro ; V C Hascall ; A J DaySource :
- Glycobiology [ 0959-6658 ] ; 2001.
Descripteurs français
- KwdFr :
- Acide hyaluronique (), Acide hyaluronique (isolement et purification), Animaux, Chondroïtines sulfate (), Chromatographie d'échange d'ions (), Chromatographie en phase liquide à haute performance (), Chromatographie sur gel (), Colorants fluorescents (), Cordon ombilical (), Humains, Hyaluronoglucosaminidase (métabolisme), Masse moléculaire, Mâle, Nouveau-né, Oligosaccharides (analyse), Oligosaccharides (isolement et purification), Ovis, Spectrométrie de masse MALDI (), Séquence glucidique, Électrophorèse ().
- MESH :
- analyse : Oligosaccharides.
- isolement et purification : Acide hyaluronique, Oligosaccharides.
- métabolisme : Hyaluronoglucosaminidase.
- Acide hyaluronique, Animaux, Chondroïtines sulfate, Chromatographie d'échange d'ions, Chromatographie en phase liquide à haute performance, Chromatographie sur gel, Colorants fluorescents, Cordon ombilical, Humains, Masse moléculaire, Mâle, Nouveau-né, Ovis, Spectrométrie de masse MALDI, Séquence glucidique, Électrophorèse.
English descriptors
- KwdEn :
- Animals, Carbohydrate Sequence, Chondroitin Sulfates (chemistry), Chromatography, Gel (methods), Chromatography, High Pressure Liquid (methods), Chromatography, Ion Exchange (methods), Electrophoresis (methods), Fluorescent Dyes (chemistry), Humans, Hyaluronic Acid (chemistry), Hyaluronic Acid (isolation & purification), Hyaluronoglucosaminidase (metabolism), Infant, Newborn, Male, Molecular Weight, Oligosaccharides (analysis), Oligosaccharides (isolation & purification), Sheep, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization (methods), Umbilical Cord (chemistry).
- MESH :
- chemical , analysis : Oligosaccharides.
- chemical , chemistry : Chondroitin Sulfates, Fluorescent Dyes, Hyaluronic Acid.
- chemical , isolation & purification : Hyaluronic Acid, Oligosaccharides.
- chemical , metabolism : Hyaluronoglucosaminidase.
- chemistry : Umbilical Cord.
- methods : Chromatography, Gel, Chromatography, High Pressure Liquid, Chromatography, Ion Exchange, Electrophoresis, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization.
- Animals, Carbohydrate Sequence, Humans, Infant, Newborn, Male, Molecular Weight, Sheep.
Abstract
Hyaluronan is a ubiquitous glycosaminoglycan of high molecular weight that acts as a structural component of extracellular matrices and mediates cell adhesion. There have been numerous recent reports that fragments of hyaluronan have different properties compared to the intact molecule. Though many of these results may be genuine, it is possible that some activities are due to minor components in the preparations used. Therefore, it is important that well-characterized and highly purified oligosaccharides are used in cell biological and structural studies so that erroneous results are avoided. We present methods for the purification of hyaluronan oligomers of defined size using size exclusion and anion-exchange chromatography following digestion of hyaluronan with testicular hyaluronidase. These preparations were characterized by a combination of electrospray ionization mass spectrometry, matrix-assisted laser desorption/ionization mass spectrometry with time-of-flight analysis, and fluorophore-assisted carbohydrate electrophoresis. Hyaluronan oligomers ranging from tetrasaccharides to 34-mers were separated. The 4- to 16-mers were shown to be homogeneous with regard to length but did contain varying amounts of chondroitin sulfate. This contaminant could have been minimized if digestion had been performed with medical-grade hyaluronan rather than the relatively impure starting material used here. The 18- to 34-mer preparations were mixtures of oligosaccharides of different lengths (e.g., the latter contained 87% 34-mer, 10% 32-mer, and 3% 30-mer) but were free of detectable chondroitin sulfate. In addition to oligomers with even numbers of sugar rings, novel 5- and 7-mers with terminal glucuronic acid residues were identified.
DOI: 10.1093/glycob/11.12.1025
PubMed: 11805075
Affiliations:
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Calabro</name></author><author><name sortKey="Hascall, V C" sort="Hascall, V C" uniqKey="Hascall V" first="V C" last="Hascall">V C Hascall</name></author><author><name sortKey="Day, A J" sort="Day, A J" uniqKey="Day A" first="A J" last="Day">A J Day</name></author></analytic><series><title level="j">Glycobiology</title><idno type="ISSN">0959-6658</idno><imprint><date when="2001" type="published">2001</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Carbohydrate Sequence</term><term>Chondroitin Sulfates (chemistry)</term><term>Chromatography, Gel (methods)</term><term>Chromatography, High Pressure Liquid (methods)</term><term>Chromatography, Ion Exchange (methods)</term><term>Electrophoresis (methods)</term><term>Fluorescent Dyes (chemistry)</term><term>Humans</term><term>Hyaluronic Acid (chemistry)</term><term>Hyaluronic Acid (isolation & purification)</term><term>Hyaluronoglucosaminidase (metabolism)</term><term>Infant, Newborn</term><term>Male</term><term>Molecular Weight</term><term>Oligosaccharides (analysis)</term><term>Oligosaccharides (isolation & purification)</term><term>Sheep</term><term>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization (methods)</term><term>Umbilical Cord (chemistry)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Acide hyaluronique ()</term><term>Acide hyaluronique (isolement et purification)</term><term>Animaux</term><term>Chondroïtines sulfate ()</term><term>Chromatographie d'échange d'ions ()</term><term>Chromatographie en phase liquide à haute performance ()</term><term>Chromatographie sur gel ()</term><term>Colorants fluorescents ()</term><term>Cordon ombilical ()</term><term>Humains</term><term>Hyaluronoglucosaminidase (métabolisme)</term><term>Masse moléculaire</term><term>Mâle</term><term>Nouveau-né</term><term>Oligosaccharides (analyse)</term><term>Oligosaccharides (isolement et purification)</term><term>Ovis</term><term>Spectrométrie de masse MALDI ()</term><term>Séquence glucidique</term><term>Électrophorèse ()</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Oligosaccharides</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Chondroitin Sulfates</term><term>Fluorescent Dyes</term><term>Hyaluronic Acid</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Hyaluronic Acid</term><term>Oligosaccharides</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Hyaluronoglucosaminidase</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>Oligosaccharides</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Umbilical Cord</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Acide hyaluronique</term><term>Oligosaccharides</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Chromatography, Gel</term><term>Chromatography, High Pressure Liquid</term><term>Chromatography, Ion Exchange</term><term>Electrophoresis</term><term>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Hyaluronoglucosaminidase</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Carbohydrate Sequence</term><term>Humans</term><term>Infant, Newborn</term><term>Male</term><term>Molecular Weight</term><term>Sheep</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Acide hyaluronique</term><term>Animaux</term><term>Chondroïtines sulfate</term><term>Chromatographie d'échange d'ions</term><term>Chromatographie en phase liquide à haute performance</term><term>Chromatographie sur gel</term><term>Colorants fluorescents</term><term>Cordon ombilical</term><term>Humains</term><term>Masse moléculaire</term><term>Mâle</term><term>Nouveau-né</term><term>Ovis</term><term>Spectrométrie de masse MALDI</term><term>Séquence glucidique</term><term>Électrophorèse</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Hyaluronan is a ubiquitous glycosaminoglycan of high molecular weight that acts as a structural component of extracellular matrices and mediates cell adhesion. There have been numerous recent reports that fragments of hyaluronan have different properties compared to the intact molecule. Though many of these results may be genuine, it is possible that some activities are due to minor components in the preparations used. Therefore, it is important that well-characterized and highly purified oligosaccharides are used in cell biological and structural studies so that erroneous results are avoided. We present methods for the purification of hyaluronan oligomers of defined size using size exclusion and anion-exchange chromatography following digestion of hyaluronan with testicular hyaluronidase. These preparations were characterized by a combination of electrospray ionization mass spectrometry, matrix-assisted laser desorption/ionization mass spectrometry with time-of-flight analysis, and fluorophore-assisted carbohydrate electrophoresis. Hyaluronan oligomers ranging from tetrasaccharides to 34-mers were separated. The 4- to 16-mers were shown to be homogeneous with regard to length but did contain varying amounts of chondroitin sulfate. This contaminant could have been minimized if digestion had been performed with medical-grade hyaluronan rather than the relatively impure starting material used here. The 18- to 34-mer preparations were mixtures of oligosaccharides of different lengths (e.g., the latter contained 87% 34-mer, 10% 32-mer, and 3% 30-mer) but were free of detectable chondroitin sulfate. In addition to oligomers with even numbers of sugar rings, novel 5- and 7-mers with terminal glucuronic acid residues were identified.</div></front></TEI><affiliations><list><country><li>Royaume-Uni</li></country><region><li>Angleterre</li><li>Oxfordshire</li></region><settlement><li>Oxford</li></settlement><orgName><li>Université d'Oxford</li></orgName></list><tree><noCountry><name sortKey="Aplin, R T" sort="Aplin, R T" uniqKey="Aplin R" first="R T" last="Aplin">R T Aplin</name><name sortKey="Calabro, A" sort="Calabro, A" uniqKey="Calabro A" first="A" last="Calabro">A. Calabro</name><name sortKey="Day, A J" sort="Day, A J" uniqKey="Day A" first="A J" last="Day">A J Day</name><name sortKey="Hascall, V C" sort="Hascall, V C" uniqKey="Hascall V" first="V C" last="Hascall">V C Hascall</name></noCountry><country name="Royaume-Uni"><region name="Angleterre"><name sortKey="Mahoney, D J" sort="Mahoney, D J" uniqKey="Mahoney D" first="D J" last="Mahoney">D J Mahoney</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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