Gene Ontology annotation through association of InterPro records with GO terms
Annotation inferences using phylogenetic trees
Gene Ontology annotation based on UniProtKB/Swiss-Prot Subcellular Location vocabulary mapping, accompanied by conservative changes to GO terms applied by UniProt
Combined Automated Annotation using Multiple IEA Methods
A mutation in the human ortholog of the Saccharomyces cerevisiae ALG6 gene causes carbohydrate-deficient glycoprotein syndrome type-Ic.
Analysis of multiple mutations in the hALG6 gene in a patient with congenital disorder of glycosylation Ic.
Defining the membrane proteome of NK cells.
Reduced expression of the oligosaccharyltransferase exacerbates protein hypoglycosylation in cells lacking the fully assembled oligosaccharide donor.
Dual proteome-scale networks reveal cell-specific remodeling of the human interactome.
Biosynthesis of the N-glycan precursor (dolichol lipid-linked oligosaccharide, LLO) and transfer to a nascent protein
Addition of the first glucose to the N-glycan precursor by ALG6
Defective ALG6 does not add glucose to the N-glycan precursor
UniProtKB Q9Y672 (ALG6_HUMAN) record
Multi-allelic origin of congenital disorder of glycosylation (CDG)-Ic.
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Human CDG-Ic fibroblasts accumulate the unglucosylated Man9GlcNAc2 lipid-linked precursor; patient-allele defects were assessed by yeast rescue.
"The detrimental effect of these mutations on ALG6 activity was confirmed by complementation of alg6 yeast mutants."
Structure and mechanism of the ER-based glucosyltransferase ALG6.
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Purified yeast ALG6 transfers glucose to a synthetic short-dolichol Man9GlcNAc2 acceptor; upstream ALG3, ALG9 and ALG12 prepare that acceptor.
"Incubation of ALG6 with Dol25-P-Glc and Dol25-PP-GlcNAc2Man9 led to complete conversion to Dol25-PP-GlcNAc2Man9Glc (Fig. 1b)."
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The structural and biochemical construct is Saccharomyces cerevisiae ALG6 Q12001, expressed by the Sf9 procedure despite human codon optimization.
"Synthetic genes (GenScript) of Flag-EYFP-PreScission-full length ALG6 (UniProtKB: Q12001), ALG9 (UniProtKB: P53868) or ALG12 (UniProtKB: P53730) from S. cerevisiae were optimized for expression in Homo sapiens (GeneArt, Thermo Fisher)."
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Yeast Asp69 mutagenesis supports a catalytic-base role; residue numbering and the measured topology are not asserted as human measurements.
"Finally, mutating Asp69 to an alanine abolished ALG6 function, and mutating it to an asparagine strongly reduced activity."