Substrate specificity of α2↔3‐sialyltransferases in ganglioside biosynthesis of rat liver golgi<sup>*</sup>

Iber, Heinrich; Echten, G van; Sandhoff, Konrad

doi:10.1111/j.1432-1033.1991.tb15683.x

Cited by 52 publications

(25 citation statements)

References 22 publications

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“…Asialo-fetuin was also examined. None of the acceptors examined showed significant levels of [ 14 C]NeuAc incorporation except lactosylceramide, indicating that this sialyltransferase is almost specific for the synthesis of GM3 as reported by Iber et al (25). Only GA1 and GA2 showed minimal levels of incorporation.…”

Section: Resultsmentioning

confidence: 67%

Expression Cloning of Mouse cDNA of CMP-NeuAc:Lactosylceramide α2,3-Sialyltransferase, an Enzyme That Initiates the Synthesis of Gangliosides

Fukumoto

Miyazaki

Goto

et al. 1999

Journal of Biological Chemistry

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Expression cloning of a cDNA for the ␣2,3-sialyltransferase (GM3 synthase) (EC 2.4.99.-) gene was performed using a GM3-lacking mouse fibroblast line L cell and anti-GM3 monoclonal antibody. Plasmids from a cDNA library generated with poly(A) ؉ RNA of a mouse fibrosarcoma line CMS5j and pdl3027 (polyoma T antigen) were co-transfected into L cells. The isolated cDNA clone pM3T-7 predicted a type II membrane protein with 13 amino acids of cytoplasmic domain, 17 amino acids of transmembrane region, and a large catalytic domain with 329 amino acids. Introduction of the cDNA clone into L cells resulted in the neo-synthesis of GM3 and high activity of ␣2,3-sialyltransferase. Among glycosphingolipids, only lactosylceramide showed significant activity as an acceptor, indicating that this gene product is a sialyltransferase specific for the synthesis of GM3. An amino acid sequence deduced from the cloned cDNA showed the typical sialyl motif with common features among ␣2,3-sialyltransferases. Among various mouse tissues, brain, liver, and testis showed relatively high expression of a 2.3-kilobase mRNA, whereas all tissues, more or less, expressed this gene.

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Section: Resultsmentioning

confidence: 67%

Expression Cloning of Mouse cDNA of CMP-NeuAc:Lactosylceramide α2,3-Sialyltransferase, an Enzyme That Initiates the Synthesis of Gangliosides

Fukumoto

Miyazaki

Goto

et al. 1999

Journal of Biological Chemistry

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“…41. In a total volume of 20 l, 200 M long chain LacCer, 0.05% Triton CF 54, 50 mM Hepes/KOH, pH 6.5, 100 mM KOAc, 5 mM Mg(OAc) 2 , 10 mM MnCl 2 , 1 mM […”

Section: Assays Of Ganglioside Biosynthesis With Long Chain Substratementioning

confidence: 99%

“…This has prompted us to investigate the localization within the Golgi of this enzyme and to reinvestigate the topography of various additional glycosyltransferases involved in GSL biosynthesis using sucrose density gradient centrifugation. To this end, the conditions for assaying enzyme activities in partially separated Golgi subfractions (25,26,41) were optimized. Using a truncated analogue of ceramide with only eight carbon atoms in both its sphingosine and fatty acid moieties (C 8 C 8 -ceramide), transferase activities can be measured in the absence of detergent, because the truncated substrates readily permeate mem-branes (39,42).…”

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confidence: 99%

Functional Organization of the Golgi Apparatus in Glycosphingolipid Biosynthesis

Lannert¹,

Gorgas²,

Meißner³

et al. 1998

Journal of Biological Chemistry

119

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Biosynthesis of plasma membrane sphingolipids involves the coordinate action of enzymes localized to individual compartments of the biosynthetic secretory pathway of proteins. These stations include the endoplasmic reticulum and the Golgi apparatus. Although a precise localization of all the enzymes that synthesize glycosphingolipids has not been achieved to date, it is assumed that the sequence of events in glycosphingolipid biosynthesis resembles that in glycoprotein biosynthesis, i.e. that early reactions occur in early stations (endoplasmic reticulum and cis/medial Golgi) of the pathway, and late reactions occur in late stations (trans Golgi/trans Golgi network).Using truncated analogues of ceramide and glucosylceramide that allow measurement of enzyme activities in intact membrane fractions, we have reinvestigated the localization of individual enzymes involved in glycosphingolipid biosynthesis and for the first time studied the localization of lactosylceramide synthase after partial separation of Golgi membranes as previously described (Trinchera, M., and Ghidoni, R. (1989) J. Biol. Chem. 264, 15766 -15769). Here, we show that the reactions involved in higher glycosphingolipid biosynthesis, including lactosylceramide synthesis, all reside in the lumen of the late Golgi compartments from rat liver.

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“…The kinetic data also suggested the existence of another species of sialyltransferase, which synthesizes GM3 from lactosylceramide [24]. GaZp3GalNAc-u3STMB also synthesizes GMlb, GDla and GTlb from asialo-GMI, GMla and GDlb, respectively.…”

Section: Discussionmentioning

confidence: 91%

Molecular cloning and expression of Galβ1,3GalNAcα2,3‐sialyltransferase from mouse brain

Lee

Kurosawa

Hamamoto

et al. 1993

European Journal of Biochemistry

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DNA clones encoding β‐galactoside α2,3‐sialyltransferase have been isolated from mouse brain cDNA libraries using sequence information obtained from the conserved amino acid sequence of the previously cloned enzymes. The cDNA sequence revealed an open reading frame coding for 337 amino acids, and the deduced amono acid sequence showed 80% identity with that of porcine submaxillary gland Galβ1,3GalNAc α2,3‐sialyltransferase. The primary structure of this enzyme suggested a putative domain structure, like that in other glycosyltransferases, consisting of a short NH2‐terminal cytoplasmic domain, a signal‐membrane anchor domain, a proteolytically sensitive stem region, and a large COOH‐terminal active domain. The identity of this enzyme was confirmed by construction of a recombinant sialyltransferase in which the NH2‐terminal part including the cytoplasmic tail, signal‐anchor domain and stem region was replaced with an immuno‐globulin signal sequence. The expression of this recombinant in COS‐7 cells resulted in secretion of a catalytically active and soluble form of the enzyme into the medium. This enzyme exhibited the transferase activity toward only the disaccharide moiety of Galβ1,3GalNAc of glycoproteins and glycolipids, no significant activity being detected for the other substrates tested.

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Substrate specificity of α2↔3‐sialyltransferases in ganglioside biosynthesis of rat liver golgi^*

Cited by 52 publications

References 22 publications

Expression Cloning of Mouse cDNA of CMP-NeuAc:Lactosylceramide α2,3-Sialyltransferase, an Enzyme That Initiates the Synthesis of Gangliosides

Expression Cloning of Mouse cDNA of CMP-NeuAc:Lactosylceramide α2,3-Sialyltransferase, an Enzyme That Initiates the Synthesis of Gangliosides

Functional Organization of the Golgi Apparatus in Glycosphingolipid Biosynthesis

Molecular cloning and expression of Galβ1,3GalNAcα2,3‐sialyltransferase from mouse brain

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Substrate specificity of α2↔3‐sialyltransferases in ganglioside biosynthesis of rat liver golgi*

Cited by 52 publications

References 22 publications

Expression Cloning of Mouse cDNA of CMP-NeuAc:Lactosylceramide α2,3-Sialyltransferase, an Enzyme That Initiates the Synthesis of Gangliosides

Expression Cloning of Mouse cDNA of CMP-NeuAc:Lactosylceramide α2,3-Sialyltransferase, an Enzyme That Initiates the Synthesis of Gangliosides

Functional Organization of the Golgi Apparatus in Glycosphingolipid Biosynthesis

Molecular cloning and expression of Galβ1,3GalNAcα2,3‐sialyltransferase from mouse brain

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Substrate specificity of α2↔3‐sialyltransferases in ganglioside biosynthesis of rat liver golgi^*