A quantitative mannose 6-phosphate receptor-based in vitro assay for recombinant human N-acetylgalactosamine-4-sulfatase.
Identifieur interne : 004961 ( Main/Exploration ); précédent : 004960; suivant : 004962A quantitative mannose 6-phosphate receptor-based in vitro assay for recombinant human N-acetylgalactosamine-4-sulfatase.
Auteurs : M. Kleinig [Australie] ; J. CoxSource :
- Analytical biochemistry [ 0003-2697 ] ; 1998.
Descripteurs français
- KwdFr :
- Adsorption (MeSH), Animaux (MeSH), Bovins (MeSH), Chondro-4-sulfatase (analyse), Chondro-4-sulfatase (métabolisme), Cinétique (MeSH), Fibroblastes (MeSH), Fixation compétitive (MeSH), Humains (MeSH), Hymécromone (analogues et dérivés), Hymécromone (métabolisme), Lignée cellulaire (MeSH), Protéines recombinantes (analyse), Protéines recombinantes (métabolisme), Récepteur IGF de type 2 (métabolisme), Spectrométrie de fluorescence (méthodes).
- MESH :
- analogues et dérivés : Hymécromone.
- analyse : Chondro-4-sulfatase, Protéines recombinantes.
- métabolisme : Chondro-4-sulfatase, Hymécromone, Protéines recombinantes, Récepteur IGF de type 2.
- méthodes : Spectrométrie de fluorescence.
- Adsorption, Animaux, Bovins, Cinétique, Fibroblastes, Fixation compétitive, Humains, Lignée cellulaire.
English descriptors
- KwdEn :
- Adsorption (MeSH), Animals (MeSH), Binding, Competitive (MeSH), Cattle (MeSH), Cell Line (MeSH), Chondro-4-Sulfatase (analysis), Chondro-4-Sulfatase (metabolism), Fibroblasts (MeSH), Humans (MeSH), Hymecromone (analogs & derivatives), Hymecromone (metabolism), Kinetics (MeSH), Receptor, IGF Type 2 (metabolism), Recombinant Proteins (analysis), Recombinant Proteins (metabolism), Spectrometry, Fluorescence (methods).
- MESH :
- chemical , analogs & derivatives : Hymecromone.
- chemical , analysis : Chondro-4-Sulfatase, Recombinant Proteins.
- chemical , metabolism : Chondro-4-Sulfatase, Hymecromone, Receptor, IGF Type 2, Recombinant Proteins.
- methods : Spectrometry, Fluorescence.
- Adsorption, Animals, Binding, Competitive, Cattle, Cell Line, Fibroblasts, Humans, Kinetics.
Abstract
An assay was developed, using two similar formats, to simultaneously measure both the lysosomal targeting receptor binding and enzyme activity of the recombinant human enzyme N-acetylgalactosamine-4-sulfatase. This assay also has potential application for all phosphorylated lysosomal enzymes that contain mannose-6-phosphate residues. The receptor was either purified from fetal bovine sera then adsorbed, or produced in situ by growing and fixing diploid human fibroblast-like cells, to a solid phase. The enzyme substrate was 4-methylumbelliferyl sulfate which fluoresces after cleavage of the sulfate moiety. Both the precursor and mature forms of the recombinant enzyme were used to demonstrate the specificity and usefulness of the assay. The assay is rapid and sensitive and has a wide dynamic range. Association between the receptor and the mannose-6-phosphate residues was abrogated in the presence of a competitive inhibitor, mannose 6-phosphate. However, partial activity was still measured when the mature enzyme was incubated in the presence of mannose 6-phosphate when using the fixed fibroblast format. This would indicate that the recombinant enzymes contain at least one terminal sugar moiety other than mannose 6-phosphate which can recognize receptors on the surface of human fibroblast-like cells. Other possible applications of this assay are also discussed.
DOI: 10.1006/abio.1998.2699
PubMed: 9657868
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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Cox</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="1998">1998</date><idno type="RBID">pubmed:9657868</idno><idno type="pmid">9657868</idno><idno type="doi">10.1006/abio.1998.2699</idno><idno type="wicri:Area/Main/Corpus">004956</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">004956</idno><idno type="wicri:Area/Main/Curation">004956</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Curation">004956</idno><idno type="wicri:Area/Main/Exploration">004956</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">A quantitative mannose 6-phosphate receptor-based in vitro assay for recombinant human N-acetylgalactosamine-4-sulfatase.</title><author><name sortKey="Kleinig, M" sort="Kleinig, M" uniqKey="Kleinig M" first="M" last="Kleinig">M. Kleinig</name><affiliation wicri:level="1"><nlm:affiliation>Research and Development Division, CSL Limited, 45 Poplar Road, Parkville, Victoria, 3052, Australia. mkleinig@csl.com.au</nlm:affiliation><country xml:lang="fr">Australie</country><wicri:regionArea>Research and Development Division, CSL Limited, 45 Poplar Road, Parkville, Victoria, 3052</wicri:regionArea><wicri:noRegion>3052</wicri:noRegion></affiliation></author><author><name sortKey="Cox, J" sort="Cox, J" uniqKey="Cox J" first="J" last="Cox">J. Cox</name></author></analytic><series><title level="j">Analytical biochemistry</title><idno type="ISSN">0003-2697</idno><imprint><date when="1998" type="published">1998</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Adsorption (MeSH)</term><term>Animals (MeSH)</term><term>Binding, Competitive (MeSH)</term><term>Cattle (MeSH)</term><term>Cell Line (MeSH)</term><term>Chondro-4-Sulfatase (analysis)</term><term>Chondro-4-Sulfatase (metabolism)</term><term>Fibroblasts (MeSH)</term><term>Humans (MeSH)</term><term>Hymecromone (analogs & derivatives)</term><term>Hymecromone (metabolism)</term><term>Kinetics (MeSH)</term><term>Receptor, IGF Type 2 (metabolism)</term><term>Recombinant Proteins (analysis)</term><term>Recombinant Proteins (metabolism)</term><term>Spectrometry, Fluorescence (methods)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Adsorption (MeSH)</term><term>Animaux (MeSH)</term><term>Bovins (MeSH)</term><term>Chondro-4-sulfatase (analyse)</term><term>Chondro-4-sulfatase (métabolisme)</term><term>Cinétique (MeSH)</term><term>Fibroblastes (MeSH)</term><term>Fixation compétitive (MeSH)</term><term>Humains (MeSH)</term><term>Hymécromone (analogues et dérivés)</term><term>Hymécromone (métabolisme)</term><term>Lignée cellulaire (MeSH)</term><term>Protéines recombinantes (analyse)</term><term>Protéines recombinantes (métabolisme)</term><term>Récepteur IGF de type 2 (métabolisme)</term><term>Spectrométrie de fluorescence (méthodes)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analogs & derivatives" xml:lang="en"><term>Hymecromone</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Chondro-4-Sulfatase</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Chondro-4-Sulfatase</term><term>Hymecromone</term><term>Receptor, IGF Type 2</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" qualifier="analogues et dérivés" xml:lang="fr"><term>Hymécromone</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>Chondro-4-sulfatase</term><term>Protéines recombinantes</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Spectrometry, Fluorescence</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Chondro-4-sulfatase</term><term>Hymécromone</term><term>Protéines recombinantes</term><term>Récepteur IGF de type 2</term></keywords><keywords scheme="MESH" qualifier="méthodes" xml:lang="fr"><term>Spectrométrie de fluorescence</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Adsorption</term><term>Animals</term><term>Binding, Competitive</term><term>Cattle</term><term>Cell Line</term><term>Fibroblasts</term><term>Humans</term><term>Kinetics</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Adsorption</term><term>Animaux</term><term>Bovins</term><term>Cinétique</term><term>Fibroblastes</term><term>Fixation compétitive</term><term>Humains</term><term>Lignée cellulaire</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">An assay was developed, using two similar formats, to simultaneously measure both the lysosomal targeting receptor binding and enzyme activity of the recombinant human enzyme N-acetylgalactosamine-4-sulfatase. This assay also has potential application for all phosphorylated lysosomal enzymes that contain mannose-6-phosphate residues. The receptor was either purified from fetal bovine sera then adsorbed, or produced in situ by growing and fixing diploid human fibroblast-like cells, to a solid phase. The enzyme substrate was 4-methylumbelliferyl sulfate which fluoresces after cleavage of the sulfate moiety. Both the precursor and mature forms of the recombinant enzyme were used to demonstrate the specificity and usefulness of the assay. The assay is rapid and sensitive and has a wide dynamic range. Association between the receptor and the mannose-6-phosphate residues was abrogated in the presence of a competitive inhibitor, mannose 6-phosphate. However, partial activity was still measured when the mature enzyme was incubated in the presence of mannose 6-phosphate when using the fixed fibroblast format. This would indicate that the recombinant enzymes contain at least one terminal sugar moiety other than mannose 6-phosphate which can recognize receptors on the surface of human fibroblast-like cells. Other possible applications of this assay are also discussed.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">9657868</PMID><DateCompleted><Year>1998</Year><Month>08</Month><Day>25</Day></DateCompleted><DateRevised><Year>2013</Year><Month>11</Month><Day>21</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0003-2697</ISSN><JournalIssue CitedMedium="Print"><Volume>260</Volume><Issue>2</Issue><PubDate><Year>1998</Year><Month>Jul</Month><Day>01</Day></PubDate></JournalIssue><Title>Analytical biochemistry</Title><ISOAbbreviation>Anal Biochem</ISOAbbreviation></Journal><ArticleTitle>A quantitative mannose 6-phosphate receptor-based in vitro assay for recombinant human N-acetylgalactosamine-4-sulfatase.</ArticleTitle><Pagination><MedlinePgn>128-34</MedlinePgn></Pagination><Abstract><AbstractText>An assay was developed, using two similar formats, to simultaneously measure both the lysosomal targeting receptor binding and enzyme activity of the recombinant human enzyme N-acetylgalactosamine-4-sulfatase. This assay also has potential application for all phosphorylated lysosomal enzymes that contain mannose-6-phosphate residues. The receptor was either purified from fetal bovine sera then adsorbed, or produced in situ by growing and fixing diploid human fibroblast-like cells, to a solid phase. The enzyme substrate was 4-methylumbelliferyl sulfate which fluoresces after cleavage of the sulfate moiety. Both the precursor and mature forms of the recombinant enzyme were used to demonstrate the specificity and usefulness of the assay. The assay is rapid and sensitive and has a wide dynamic range. Association between the receptor and the mannose-6-phosphate residues was abrogated in the presence of a competitive inhibitor, mannose 6-phosphate. However, partial activity was still measured when the mature enzyme was incubated in the presence of mannose 6-phosphate when using the fixed fibroblast format. This would indicate that the recombinant enzymes contain at least one terminal sugar moiety other than mannose 6-phosphate which can recognize receptors on the surface of human fibroblast-like cells. Other possible applications of this assay are also discussed.</AbstractText><CopyrightInformation>Copyright 1998 Academic Press.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Kleinig</LastName><ForeName>M</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>Research and Development Division, CSL Limited, 45 Poplar Road, Parkville, Victoria, 3052, Australia. mkleinig@csl.com.au</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Cox</LastName><ForeName>J</ForeName><Initials>J</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Anal Biochem</MedlineTA><NlmUniqueID>0370535</NlmUniqueID><ISSNLinking>0003-2697</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D017527">Receptor, IGF Type 2</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011994">Recombinant Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>25892-63-1</RegistryNumber><NameOfSubstance UI="C019492">4-methylumbelliferyl sulfate</NameOfSubstance></Chemical><Chemical><RegistryNumber>3T5NG4Q468</RegistryNumber><NameOfSubstance UI="D006923">Hymecromone</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 3.1.6.9</RegistryNumber><NameOfSubstance UI="D002803">Chondro-4-Sulfatase</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000327" MajorTopicYN="N">Adsorption</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D000818" MajorTopicYN="N">Animals</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D001667" MajorTopicYN="N">Binding, Competitive</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002417" MajorTopicYN="N">Cattle</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002460" MajorTopicYN="N">Cell Line</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002803" MajorTopicYN="N">Chondro-4-Sulfatase</DescriptorName><QualifierName UI="Q000032" MajorTopicYN="N">analysis</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D005347" MajorTopicYN="N">Fibroblasts</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006923" MajorTopicYN="N">Hymecromone</DescriptorName><QualifierName UI="Q000031" MajorTopicYN="N">analogs & derivatives</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D007700" MajorTopicYN="N">Kinetics</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D017527" MajorTopicYN="N">Receptor, IGF Type 2</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011994" MajorTopicYN="N">Recombinant Proteins</DescriptorName><QualifierName UI="Q000032" MajorTopicYN="N">analysis</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D013050" MajorTopicYN="N">Spectrometry, Fluorescence</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="N">methods</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>1998</Year><Month>7</Month><Day>11</Day></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>1998</Year><Month>7</Month><Day>11</Day><Hour>0</Hour><Minute>1</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>1998</Year><Month>7</Month><Day>11</Day><Hour>0</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">9657868</ArticleId><ArticleId IdType="pii">S0003-2697(98)92699-9</ArticleId><ArticleId IdType="doi">10.1006/abio.1998.2699</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Australie</li></country></list><tree><noCountry><name sortKey="Cox, J" sort="Cox, J" uniqKey="Cox J" first="J" last="Cox">J. Cox</name></noCountry><country name="Australie"><noRegion><name sortKey="Kleinig, M" sort="Kleinig, M" uniqKey="Kleinig M" first="M" last="Kleinig">M. Kleinig</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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