2.4.1.90 C134S complete loss of activity 489552 2.4.1.90 C342S 33fold increase in the apparent Km-value for UDPgalactose 489552 2.4.1.90 D254E 0.01% of the activity of the wild-type enzyme 489556 2.4.1.90 D254N 0.01% of the activity of the wild-type enzyme 489556 2.4.1.90 D315A the mutant shows about 90% of wild type activity 758173 2.4.1.90 D320A when partially activated by Mn2+ binding to the primary site, can be further activated by Co2+ or inhibited by Ca2+, an effect that is the opposite of what is observed with the wild-type enzyme 489556 2.4.1.90 D320E when partially activated by Mn2+ binding to the primary site, can be further activated by Co2+ or inhibited by Ca2+, an effect that is the opposite of what is observed with the wild-type enzyme 489556 2.4.1.90 D320N when partially activated by Mn2+ binding to the primary site, can be further activated by Co2+ or inhibited by Ca2+, an effect that is the opposite of what is observed with the wild-type enzyme 489556 2.4.1.90 DELTA1-70 crystallization of a refolded Drosophila Cd7 protein coding the catalytic domain (Cd7) sequence (residues 71-322) is not successful 719863 2.4.1.90 DELTA1-70/DELTA312-322 a DNA fragment bearing the Cd7 catalytic domain and C-terminal 11-amino acid deletion (Cd7DELTAC) sequence (residues 71-311) shows no difference in catalytic activity or folding but crystalization is not possible 719863 2.4.1.90 E317A when partially activated by Mn2+ binding to the primary site, can be further activated by Co2+ or inhibited by Ca2+, an effect that is the opposite of what is observed with the wild-type enzyme 489556 2.4.1.90 E317D when partially activated by Mn2+ binding to the primary site, can be further activated by Co2+ or inhibited by Ca2+, an effect that is the opposite of what is observed with the wild-type enzyme 489556 2.4.1.90 E317Q when partially activated by Mn2+ binding to the primary site, can be further activated by Co2+ or inhibited by Ca2+, an effect that is the opposite of what is observed with the wild-type enzyme 489556 2.4.1.90 H343A the mutant shows about 3% of wild type activity 758173 2.4.1.90 H347D in presence of Mn2+ retains 0.02% of wild-type enzyme activity, in presence of Co2+ retains 0.085% of wild-type enzyme activity 489556 2.4.1.90 H347E in presence of Mn2+ retains 0.1% of wild-type enzyme activity, in presence of Co2+ retains 0.4% of wild-type enzyme activity 489556 2.4.1.90 H347N in presence of Mn2+ retains 0.07% of wild-type enzyme activity, in presence of Co2+ retains 0.36% of wild-type enzyme activity 489556 2.4.1.90 H347Q in presence of Mn2+ retains 0.28% of wild-type enzyme activity, in presence of Co2+ retains 1.21% of wild-type enzyme activity 489556 2.4.1.90 I285Y site-directed mutagenesis, the mutation converts the betaGALNAcT1 enzyme into an efficient beta4Gal-T1, the N-acetylgalactosaminyltransferase activity is reduced by nearly 1000fold, while the galactosyltransferase activity is enhanced by 80fold 675421 2.4.1.90 M340E the mutant shows about 85% of wild type activity 758173 2.4.1.90 M340H the mutant shows about 10% of wild type activity 758173 2.4.1.90 M344A in presence of Mn2+ retains 54.5% of wild-type enzyme activity, in presence of Co2+ retains 6.15% of wild-type enzyme activity 489556 2.4.1.90 M344A site-directed mutagenesis, altered metal ion specificity compared to the wild-type enzyme 658032 2.4.1.90 M344E site-directed mutagenesis, altered metal ion specificity compared to the wild-type enzyme 658032 2.4.1.90 M344H mutant prefers Mg2+ instead of Mn2+ which is preferred by the wild-type enzyme, with Mn2+ the mutant is arrested in the closed inactive conformation 658502 2.4.1.90 M344H site-directed mutagenesis, the mutant shows altered conformational changes upon binding of Mn2+ and substrates or substrate analogues compared to the wild-type enzyme, it forms the closed conformation with bound Mn2+ and UDP-hexanolamine, loss of 98% of Mn2+ binding activity, increased activity with Mg2+ 658032 2.4.1.90 M344Q in presence of Mn2+ retains 15.37% of wild-type enzyme activity, in presence of Co2+ retains 31.08% of wild-type enzyme activity 489556 2.4.1.90 M344Q site-directed mutagenesis, altered metal ion specificity compared to the wild-type enzyme 658032 2.4.1.90 M344S site-directed mutagenesis, altered metal ion specificity compared to the wild-type enzyme 658032 2.4.1.90 additional information a construct bearing the N-terminal peptides from bovine Cd1 (residues 143–175 (P1)) fused with peptide with the Cd7DELTAC protein (last 11 amino acids deleted) shows no impact on the catalytic activity nor the folding efficiency. This construct is used for successful crystallization 719863 2.4.1.90 additional information decreasing the expression of beta1,4GalT II using RNA interference inhibits p53-mediated HeLa cell apoptosis induced by adriamycin 687455 2.4.1.90 additional information mutations of Asp318 and Asp319 abolish enzyme activity 489556 2.4.1.90 additional information N-terminal truncated forms of the enzyme between residues 1-129, do not show any significant difference in the apparent Km-values towards N-acetylglucosamine or linear oligosaccharide acceptors, e.g. for chitobiose and chitotriose, or for the nucleotide donor UDPgalactose. The binding behaviour of N-terminal and C-terminal fragments of the enzyme towards the N-acetylglucosamine-agarose and UDP-agarose columns differ, the former binds preferentially to the N-acetylglucosamine-columns, while the latter binds to UDP-agarose columns via Mn2+ 489552 2.4.1.90 additional information the enzyme transfected 7721 hepatocarcinoma cell line is more susceptible to cycloheximide-induced apoptosis than the wild-type cells, cells show increased enzyme activity and expression of Galbeta1,4GlcNAc groups on the cell surface 659925 2.4.1.90 W314A site-directed mutagenesis, mutant shows highly reduced activity, i.e. 0.6% of wild-type galactosyltransferase activity, substrate binding, and reduced binding to the effector alpha-lactalbumin as well as reduced susceptibility to cleavage by proteases Glu-C and Lys-C compared to the wild-type enzyme, overview 659700 2.4.1.90 Y268G/W294G inactive 757209 2.4.1.90 Y289I mutant shows high activity with UDP-GalNAc in contrary to the wild-type enzyme 658502 2.4.1.90 Y289I mutation enhances GalNAc-transferase activity. Km for GlcNAc is increased compared to the wild type 489546 2.4.1.90 Y289L mutant shows high activity with UDP-GalNAc in contrary to the wild-type enzyme 658502 2.4.1.90 Y289L mutation enhances GalNAc-transferase activity. Km for GlcNAc is incereased compared to the wild type 489546 2.4.1.90 Y289L/C342T site-directed mutagenesis, the mutant is able to transfer GalNAc from the sugar donor UDP-GalNAc to the acceptor, GlcNAc, with efficiency as good as that of galactose from UDP-Gal, in contrast to the wild-type enzyme, mutant substrate specificity with different donor substrate and oligosaccharides as acceptor substrates, mass spectrometry product analysis, overview, the C342T mutation does not alter enzyme activity, but increases the enzyme stability at room temperature 672454 2.4.1.90 Y289N mutation enhances GalNAc-transferase activity. Km for GlcNAc is increased compared to the wild type 489546