ArticleThe Biochemical journal1995
The effects of the site-directed removal of N-glycosylation sites from beta-1,4-N-acetylgalactosaminyltransferase on its function.
Article in The Biochemical journal, 1995. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed.
- Characterization of Human Recombinant β1,4-GalNAc-Transferase B4GALNT1 and Inhibition by Selected Compounds.Molecules (Basel, Switzerland) · 2025Article
- Sugar Auxiliary Group Assisted Diversity-Oriented Enzymatic Modular Synthesis of 0-Series Ganglioside Glycans.Angewandte Chemie (International ed. in English) · 2025Article
- Article
- Effect of3 Biotech · 2022Article
- N-glycosylation of the human β1,4-galactosyltransferase 4 is crucial for its activity and Golgi localization.Glycoconjugate journal · 2020Article
- Article
- Protein Quality Control in the Endoplasmic Reticulum.The protein journal · 2019Review
- Trans-activity of plasma membrane-associated ganglioside sialyltransferase in mammalian cells.The Journal of biological chemistry · 2011Article
- Role of glycosylation in conformational stability, activity, macromolecular interaction and immunogenicity of recombinant human factor VIII.The AAPS journal · 2009Article
- Role of the linker region in the expression of Rhizopus oryzae glucoamylase.BMC biochemistry · 2007Article
- N-linked oligosaccharides are required to produce and stabilize the active form of chondroitin 4-sulphotransferase-1.The Biochemical journal · 2005Article
- The STT3a subunit isoform of the Arabidopsis oligosaccharyltransferase controls adaptive responses to salt/osmotic stress.The Plant cell · 2003Article
- N-glycosylation is required for full enzymic activity of the murine galactosylceramide sulphotransferase.The Biochemical journal · 2002Article
- Understanding the stepwise synthesis of glycolipids.Neurochemical research · 2002Review
- Glycosylation of the N-terminal potential N-glycosylation sites in the human alpha1,3-fucosyltransferase V and -VI (hFucTV and -VI).Glycoconjugate journal · 2000Article
- GM1 synthase depends on N-glycosylation for enzyme activity and trafficking to the Golgi complex.Neurochemical research · 2000Article
- Expression of the Schwanniomyces occidentalis SWA2 amylase in Saccharomyces cerevisiae: role of N-glycosylation on activity, stability and secretion.The Biochemical journal · 1998Article
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The amino acid sequence deduced from the cloned human cDNA of beta-1,4-N-acetylgalactosaminyltransferase (GalNAc-T; EC 2.4.1.92) gene predicted three potential sites for N-linked glycosylation. Although many glycosyltransferases isolated contain from 2 to 6 N-glycosylation sites, their significance has not been adequately demonstrated. To clarify the roles of N-glycosylation in GalNAc-T function, we generated a series of mutant cDNAs, in which some or all of the glycosylation recognition sites were eliminated by polymerase chain reaction (PCR)-mediated site-directed mutagenesis. Using transcription/translation in vitro, we confirmed that all potential N-glycosylation sites could be used. Although cell lines transfected with mutant cDNAs showed equivalent levels of GalNAc beta 1-->4(NeuAc alpha 2-->3)Gal beta 1-->4Glc-Cer (GM2) to that of the wild-type, the extracts from mutant cDNA transfectants demonstrated lower enzyme activity than in the wild-type. The decrease in enzyme activity was more evident as the number of deglycosylated sites increased, with about 90% decrease in a totally deglycosylated mutant. The enzyme kinetics analysis revealed no significant change of Km among wild-type and mutant cDNA products. The intracellular localization of GalNAc-T expressed in transfectants with wild-type or mutant cDNAs also showed a similar perinuclear pattern (Golgi pattern). These results suggest that N-linked carbohydrates on GalNAc-T are required for regulating the stability of the enzyme structure.
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