Dolichyl-phosphomannose synthase from the Archae Thermoplasma acidophilum

Dolichyl-phosphomannose synthase from the Archae Thermoplasma acidophilum Archae (formerly Archaebacteria) comprise an entire kingdom of organisms placed halfway between prokaryotes and eucaryotes in evolution. This class of organisms lacks murein cell wall and is devoid of organelles, yet Archae synthesize and export N-linked and O-linked glycoproteins utitiring only the plasma membrane. Study of glycosylation systems in Archae is extremely interesting because the plasma membrane must perform many functions normally carried out by the endoplasmic reticulum and Golgi in eucaryotes. This report represents the first glycosyl transferase system enzyme demonstrated from archae showing a functional relationship with homologous eucaryotic enzymes. Archae dolichyl-phosphoryl-mannose synthase was purified 1070-fold from Thermoplasma acidophilum by column chromatography on Sephacryl S-200, Cibacron blue 3GA-agarose, Octyl-Sepharose, and hydroxylapatite in the presence of 0.2% polioxyethylene 9 lauryl ether. The enzyme activity was stimulated by MgCl 2 (20 mM optimum) and exhibited a pH optimum at 6.0. Although the native polyisoprenoid has not been isolated or characterized, the enzyme prefers dolichyl phosphate (dol-P) to C 55 polyisoprenol as an acceptor, and the K m value for dol-P was calculated to be 2.6 μM. Amphomycin, an inhibitor of dol-P-Man synthase, blocked mannosyl transfer to the endogenous lipids, proteins, and to dol-P; 100 μg/ml amphomycin inhibited 97% of mannosyl transfer to dol-P, and 50% to endogenous acceptors, indicating direct transfer from GDP-mannose to some intermediates or final structures. The size range of ( 3 H)an-oligosaccharides from acid-labile manno-lipid product was from dp 1 to 4. dol-P Man synthase activity could be correlated directly with a 42 kDa band on SDS/polyacrylamide gel electrophoresis. Key words Archae dol-P-Man synthase Thermoplasma acidophilum © Oxford University Press « Previous | Next Article » Table of Contents This Article Glycobiology (1996) 6 (8): 811-816. doi: 10.1093/glycob/6.8.811 » Abstract Free Full Text (PDF) Free Classifications Original Article Services Article metrics Alert me when cited Alert me if corrected Find similar articles Similar articles in Web of Science Similar articles in PubMed Add to my archive Download citation Request Permissions Disclaimer Citing Articles Load citing article information Citing articles via CrossRef Citing articles via Scopus Citing articles via Web of Science Citing articles via Google Scholar Google Scholar Articles by Zhu, B. C. Articles by Laine, R. A. Search for related content PubMed PubMed citation Articles by Zhu, B. C. Articles by Laine, R. A. Related Content Load related web page information Share Email this article CiteULike Delicious Facebook Google+ Mendeley Twitter What's this? Search this journal: Advanced » Current Issue November 2015 25 (11) Alert me to new issues The Journal Submit now! About this journal Rights & Permissions Manuscript submission & review Dispatch date of the next issue This journal is a member of the Committee on Publication Ethics (COPE) We are mobile – find out more Journals Career Network Glycoscience resources Consortium for Functional Glycomics The Official Journal of The Society for Glycobiology Impact factor: 3.147 5-Yr impact factor: 3.212 Editor-in-Chief Robert S. Haltiwanger View full editorial board For Authors Instructions to authors Self-archiving policy Online submission Open access options for authors - visit Oxford Open This journal enables compliance with the NIH Public Access Policy Alerting Services Email table of contents Email Advance Access CiteTrack XML RSS feed Corporate Services Advertising sales Classified Advertising Reprints Supplements var taxonomies = ("SCI01000"); Most Most Read Protein glycosylation: nature, distribution, enzymatic formation, and disease implications of glycopeptide bonds A "Glyconutrient Sham" Formation of the glycan chains in the synthesis of bacterial peptidoglycan Fucose: biosynthesis and biological function in mammals Optimal and consistent protein glycosylation in mammalian cell culture » View all Most Read articles Most Cited Biological roles of oligosaccharides: all of the theories are correct Prediction, conservation analysis, and structural characterization of mammalian mucin-type O-glycosylation sites An evolving view of the eukaryotic oligosaccharyltransferase Evolutionary considerations in relating oligosaccharide diversity to biological function Glycosidases of the asparagine-linked oligosaccharide processing pathway » View all Most Cited articles Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department. Online ISSN 1460-2423 - Print ISSN 0959-6658 Copyright © 2015 Oxford University Press Oxford Journals Oxford University Press Site Map Privacy Policy Cookie Policy Legal Notices Frequently Asked Questions Other Oxford University Press sites: Oxford University Press Oxford Journals China Oxford Journals Japan Academic & Professional books Children's & Schools Books Dictionaries & Reference Dictionary of National Biography Digital Reference English Language Teaching Higher Education Textbooks International Education Unit Law Medicine Music Online Products & Publishing Oxford Bibliographies Online Oxford Dictionaries Online Oxford English Dictionary Oxford Language Dictionaries Online Oxford Scholarship Online Reference Rights and Permissions Resources for Retailers & Wholesalers Resources for the Healthcare Industry Very Short Introductions World's Classics function fnc_onDomLoaded() { var query_context = getQueryContext(); PF_initOIUnderbar(query_context,":QS:default","","JRN"); PF_insertOIUnderbar(0); }; if (window.addEventListener) { window.addEventListener('load', fnc_onDomLoaded, false); } else if (window.attachEvent) { window.attachEvent('onload', fnc_onDomLoaded); } var gaJsHost = (("https:" == document.location.protocol) ? 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Dolichyl-phosphomannose synthase from the Archae Thermoplasma acidophilum

Glycobiology, Volume 6 (8) – Dec 1, 1996

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Oxford University Press
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Copyright © 2015 Oxford University Press
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0959-6658
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1460-2423
DOI
10.1093/glycob/6.8.811
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Abstract

Archae (formerly Archaebacteria) comprise an entire kingdom of organisms placed halfway between prokaryotes and eucaryotes in evolution. This class of organisms lacks murein cell wall and is devoid of organelles, yet Archae synthesize and export N-linked and O-linked glycoproteins utitiring only the plasma membrane. Study of glycosylation systems in Archae is extremely interesting because the plasma membrane must perform many functions normally carried out by the endoplasmic reticulum and Golgi in eucaryotes. This report represents the first glycosyl transferase system enzyme demonstrated from archae showing a functional relationship with homologous eucaryotic enzymes. Archae dolichyl-phosphoryl-mannose synthase was purified 1070-fold from Thermoplasma acidophilum by column chromatography on Sephacryl S-200, Cibacron blue 3GA-agarose, Octyl-Sepharose, and hydroxylapatite in the presence of 0.2% polioxyethylene 9 lauryl ether. The enzyme activity was stimulated by MgCl 2 (20 mM optimum) and exhibited a pH optimum at 6.0. Although the native polyisoprenoid has not been isolated or characterized, the enzyme prefers dolichyl phosphate (dol-P) to C 55 polyisoprenol as an acceptor, and the K m value for dol-P was calculated to be 2.6 μM. Amphomycin, an inhibitor of dol-P-Man synthase, blocked mannosyl transfer to the endogenous lipids, proteins, and to dol-P; 100 μg/ml amphomycin inhibited 97% of mannosyl transfer to dol-P, and 50% to endogenous acceptors, indicating direct transfer from GDP-mannose to some intermediates or final structures. The size range of ( 3 H)an-oligosaccharides from acid-labile manno-lipid product was from dp 1 to 4. dol-P Man synthase activity could be correlated directly with a 42 kDa band on SDS/polyacrylamide gel electrophoresis. Key words Archae dol-P-Man synthase Thermoplasma acidophilum © Oxford University Press « Previous | Next Article » Table of Contents This Article Glycobiology (1996) 6 (8): 811-816. doi: 10.1093/glycob/6.8.811 » Abstract Free Full Text (PDF) Free Classifications Original Article Services Article metrics Alert me when cited Alert me if corrected Find similar articles Similar articles in Web of Science Similar articles in PubMed Add to my archive Download citation Request Permissions Disclaimer Citing Articles Load citing article information Citing articles via CrossRef Citing articles via Scopus Citing articles via Web of Science Citing articles via Google Scholar Google Scholar Articles by Zhu, B. C. Articles by Laine, R. A. Search for related content PubMed PubMed citation Articles by Zhu, B. C. Articles by Laine, R. A. Related Content Load related web page information Share Email this article CiteULike Delicious Facebook Google+ Mendeley Twitter What's this? Search this journal: Advanced » Current Issue November 2015 25 (11) Alert me to new issues The Journal Submit now! About this journal Rights & Permissions Manuscript submission & review Dispatch date of the next issue This journal is a member of the Committee on Publication Ethics (COPE) We are mobile – find out more Journals Career Network Glycoscience resources Consortium for Functional Glycomics The Official Journal of The Society for Glycobiology Impact factor: 3.147 5-Yr impact factor: 3.212 Editor-in-Chief Robert S. Haltiwanger View full editorial board For Authors Instructions to authors Self-archiving policy Online submission Open access options for authors - visit Oxford Open This journal enables compliance with the NIH Public Access Policy Alerting Services Email table of contents Email Advance Access CiteTrack XML RSS feed Corporate Services Advertising sales Classified Advertising Reprints Supplements var taxonomies = ("SCI01000"); Most Most Read Protein glycosylation: nature, distribution, enzymatic formation, and disease implications of glycopeptide bonds A "Glyconutrient Sham" Formation of the glycan chains in the synthesis of bacterial peptidoglycan Fucose: biosynthesis and biological function in mammals Optimal and consistent protein glycosylation in mammalian cell culture » View all Most Read articles Most Cited Biological roles of oligosaccharides: all of the theories are correct Prediction, conservation analysis, and structural characterization of mammalian mucin-type O-glycosylation sites An evolving view of the eukaryotic oligosaccharyltransferase Evolutionary considerations in relating oligosaccharide diversity to biological function Glycosidases of the asparagine-linked oligosaccharide processing pathway » View all Most Cited articles Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department. Online ISSN 1460-2423 - Print ISSN 0959-6658 Copyright © 2015 Oxford University Press Oxford Journals Oxford University Press Site Map Privacy Policy Cookie Policy Legal Notices Frequently Asked Questions Other Oxford University Press sites: Oxford University Press Oxford Journals China Oxford Journals Japan Academic & Professional books Children's & Schools Books Dictionaries & Reference Dictionary of National Biography Digital Reference English Language Teaching Higher Education Textbooks International Education Unit Law Medicine Music Online Products & Publishing Oxford Bibliographies Online Oxford Dictionaries Online Oxford English Dictionary Oxford Language Dictionaries Online Oxford Scholarship Online Reference Rights and Permissions Resources for Retailers & Wholesalers Resources for the Healthcare Industry Very Short Introductions World's Classics function fnc_onDomLoaded() { var query_context = getQueryContext(); PF_initOIUnderbar(query_context,":QS:default","","JRN"); PF_insertOIUnderbar(0); }; if (window.addEventListener) { window.addEventListener('load', fnc_onDomLoaded, false); } else if (window.attachEvent) { window.attachEvent('onload', fnc_onDomLoaded); } var gaJsHost = (("https:" == document.location.protocol) ? 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GlycobiologyOxford University Press

Published: Dec 1, 1996

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