2.4.1.257: GDP-Man:Man2GlcNAc2-PP-dolichol alpha-1,6-mannosyltransferase
This is an abbreviated version!
For detailed information about GDP-Man:Man2GlcNAc2-PP-dolichol alpha-1,6-mannosyltransferase, go to the full flat file.
Reaction
Synonyms
Alg2, Alg2 mannosyltransferase, Alg2 MTase, More, MTase, scAlg2
ECTree
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Engineering
Engineering on EC 2.4.1.257 - GDP-Man:Man2GlcNAc2-PP-dolichol alpha-1,6-mannosyltransferase
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E335A
E335A/E343A
significant lower level of product formation, identical to that of the E335A mutant
E343A
F337A
site-directed mutagenesis, Trx-scAlg2F337A produces 26% Man3Gn2 product compared to wild-type enzyme
G377R
site-directed mutagenesis, a temperature-sensitive alg2-1 mutant containing a single missense mutation, catalytically inactive
H336A
V62G
site-directed mutagenesis, Trx-scAlg2V62G produces 25% Man3Gn2 product compared to wild-type enzyme. The HA-tagged mutant allele (3HAscAlg2V62G) fails to complement the lethality of the alg2DELTA LSY2 when grown on 5-FOA
additional information
E335A
site-directed mutagenesis, Trx-scAlg2E335A produces only no final product and only 32% of intermediate Man2Gn2 compared to wild-type enzyme
H336A
site-directed mutagenesis, Trx-scAlg2H336A produces 8% Man3Gn2 product compared to wild-type enzyme
chemo-enzymatic synthesis of lipid-linked GlcNAc2Man5 oligosaccharides using recombinant Alg1 (from yeast, EC 2.4.1.142), Alg2, and Alg11 (from yeast, EC 2.4.1.131) proteins, chemo-enzymatic synthesis strategy to extend the glycan moiety of synthetic LLO analogues to Dol-PP-GlcNAc2Man5, method, overview
additional information
mutational analysis of Alg2 and identification of amino acids required for its activity. None of the four domains (predicted as transmembrane-spanning helices) is essential for transferase activity because truncated Alg2 variants can exert their function as long as Alg2 is associated with the endaplasmic reticulum by either its N- or C-terminal hydrophobic regions
additional information
site-directed mutagenesis of conserved EX7E motif. Trx-scAlg2E335A, mutated in the first E, has significantly decreased activity, producing no final product and only 32% of intermediate Man2Gn2. Trx-scAlg2E343A, mutated in the second E, has no detectable activity. The intervening amino acids of the EX7E are also important, though less than either E335 or E343. Trx-scAlg2H336A and Trx-scAlg2F337A produce 8% and 26% of Man3Gn2 product, respectively, compared to wild-type. Cells deleted for ALG2 are inviable, a plasmid shuffling technique is used to measure complementation. Mutant alg2 alleles display intraallelic complementation. Mutations (changed to proline) in five of the glycines (G19, G20, G256, G357, G358) result in complete loss of activity, while two of them (G17, G257) are significantly decreased