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Information on EC 2.4.1.267 - dolichyl-P-Glc:Man9GlcNAc2-PP-dolichol alpha-1,3-glucosyltransferase and Organism(s) Saccharomyces cerevisiae and UniProt Accession Q12001

for references in articles please use BRENDA:EC2.4.1.267
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EC Tree
IUBMB Comments
The successive addition of three glucose residues by EC 2.4.1.267, EC 2.4.1.265 (Dol-P-Glc:Glc1Man9GlcNAc2-PP-Dol alpha-1,3-glucosyltransferase) and EC 2.4.1.256 (Dol-P-Glc:Glc2Man9GlcNAc2-PP-Dol alpha-1,2-glucosyltransferase) represents the final stage of the lipid-linked oligosaccharide assembly.
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Saccharomyces cerevisiae
UNIPROT: Q12001
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Word Map
The taxonomic range for the selected organisms is: Saccharomyces cerevisiae
The enzyme appears in selected viruses and cellular organisms
Synonyms
halg6, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SYSTEMATIC NAME
IUBMB Comments
dolichyl beta-D-glucosyl phosphate: D-Man-alpha-(1->2)-D-Man-alpha-(1->2)-D-Man-alpha-(1->3)-[D-Man-alpha-(1->2)-D-Man-alpha-(1->3)-[D-Man-alpha-(1->2)-D-Man-alpha-(1->6)]-D-Man-alpha-(1->6)]-D-Man-beta-(1->4)-D-GlcNAc-beta-(1->4)-D-GlcNAc-diphosphodolichol alpha-1,3-mannosyltransferase
The successive addition of three glucose residues by EC 2.4.1.267, EC 2.4.1.265 (Dol-P-Glc:Glc1Man9GlcNAc2-PP-Dol alpha-1,3-glucosyltransferase) and EC 2.4.1.256 (Dol-P-Glc:Glc2Man9GlcNAc2-PP-Dol alpha-1,2-glucosyltransferase) represents the final stage of the lipid-linked oligosaccharide assembly.
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
dolichyl beta-D-glucosyl phosphate + alpha-D-Man-(1->2)-alpha-D-Man-(1->2)-alpha-D-Man-(1->3)-[alpha-D-Man-(1->2)-alpha-D-Man-(1->3)-[alpha-D-Man-(1->2)-alpha-D-Man-(1->6)]-alpha-D-Man-(1->6)]-beta-D-Man-(1->4)-beta-D-GlcNAc-(1->4)-alpha-D-GlcNAc-diphospho-Dol25
alpha-D-Glc-(1->3)-alpha-D-Man-(1->2)-alpha-D-Man-(1->2)-alpha-D-Man-(1->3)-[alpha-D-Man-(1->2)-alpha-D-Man-(1->3)-[alpha-D-Man-(1->2)-alpha-D-Man-(1->6)]-alpha-D-Man-(1->6)]-beta-D-Man-(1->4)-beta-D-GlcNAc-(1->4)-alpha-D-GlcNAc-diphospho-Dol25 + dolichyl phosphate
show the reaction diagram
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-
-
?
dolichyl beta-D-glucosyl phosphate + alpha-D-Man-(1->2)-alpha-D-Man-(1->2)-alpha-D-Man-(1->3)-[alpha-D-Man-(1->2)-alpha-D-Man-(1->3)-[alpha-D-Man-(1->2)-alpha-D-Man-(1->6)]-alpha-D-Man-(1->6)]-beta-D-Man-(1->4)-beta-D-GlcNAc-(1->4)-alpha-D-GlcNAc-diphosphodolichol
alpha-D-Glc-(1->3)-alpha-D-Man-(1->2)-alpha-D-Man-(1->2)-alpha-D-Man-(1->3)-[alpha-D-Man-(1->2)-alpha-D-Man-(1->3)-[alpha-D-Man-(1->2)-alpha-D-Man-(1->6)]-alpha-D-Man-(1->6)]-beta-D-Man-(1->4)-beta-D-GlcNAc-(1->4)-alpha-D-GlcNAc-diphosphodolichol + dolichyl phosphate
show the reaction diagram
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-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
no evidence for metal ions in the crystal structure, nor presence of a DXD motif. Reaction is metal-ion independent
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
not inhibitory: EDTA
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
alg6 mutants accumulate lipid-linked Man9GlcNAc2
physiological function
three-state mechanism, where Dol-P-Glc binds before the Man9-containing acceptor substrate, because the glucose moiety is at the bottom of the active site cavity. Donor and acceptor substrates bind sequentially and Asp69 acts as a general base that abstracts the proton of the 3-hydroxyl group of the terminal A-branch mannose of the acceptor substrate to activate it for a nucleophilic attack
malfunction
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alg6-1 mutants accumulate Man9GlcNAc2-P-P-dolichol as their largest lipid-linked oligosaccharide in vivo and in vitro. alg6-1 cells are unable to transfer glucose from dolichol phosphoglucose to the unglucosylated lipid-linked oligosaccharide
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
structure of nanodisc-reconstituted apo-ALG6 at 3 A resolution. ALG6 has 14 transmembrane helices and two long loops (EL1 and EL4) that form helices in the ER lumen. The first external loop contains the catalytically essential residue Asp69
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D307A
activity is unaffected by the mutation
D69A
loss of activity
D69N
strong reduction of activity
D99A
loss of activity
D99N
activity is unaffected by the mutation
E306A
activity is unaffected by the mutation
E379A
activity is unaffected by the mutation
H378A
strong reduction of activity
H378D
strong reduction of activity
H378E
strong reduction of activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
ALG6 is active in different detergents as well as reconstituted in lipid nanodiscs
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Reiss, G.; te Heesen, S.; Zimmerman, J.; Robbins, P.W.; Aebi, M.
Isolation of the ALG6 locus of Saccharomyces cerevisiae required for glucosylation in the N-linked glycosylation pathway
Glycobiology
6
493-498
1996
Saccharomyces cerevisiae (Q12001)
Manually annotated by BRENDA team
Runge, K.W.; Huffaker, T.C.; Robbins, P.W.
Two yeast mutations in glucosylation steps of the asparagine glycosylation pathway
J. Biol. Chem.
259
412-417
1983
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Bloch, J.S.; Pesciullesi, G.; Boilevin, J.; Nosol, K.; Irobalieva, R.N.; Darbre, T.; Aebi, M.; Kossiakoff, A.A.; Reymond, J.L.; Locher, K.P.
Structure and mechanism of the ER-based glucosyltransferase ALG6
Nature
579
443-447
2020
Saccharomyces cerevisiae (Q12001)
Manually annotated by BRENDA team