Cloned (Comment) | Organism |
---|---|
gene profut2, recombinant expression in Pichia pastoris and secretion to the medium, partially degradation of the protein | Caenorhabditis elegans |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme CePOFUT2 in complex with GDP and human TSR1, X-ray diffraction structure determination and analysis | Caenorhabditis elegans |
Protein Variants | Comment | Organism |
---|---|---|
G19Q | site-directed mutagenesis, the mutation affects both secretion and fucosylation | Caenorhabditis elegans |
G19R | site-directed mutagenesis, the mutation affects both secretion and fucosylation | Caenorhabditis elegans |
G20H | site-directed mutagenesis, the mutation affects both secretion and fucosylation | Caenorhabditis elegans |
R298K/R299K | site-directed mutagenesis, the mutant enzyme is stable against proteolysis and similarly active as the wild-type. The mutant enzymes is capable of fucosylating TSRs not only of group 1 but also of group 2, it not only recognizes and reacts with TSRs showing slightly different structures but also accepts TSRs with very low sequence identity. Residue Glu52 of mutant CePOFUT is the catalytic base | Caenorhabditis elegans |
S15D | site-directed mutagenesis, the mutant shows increased secretion, but a significant reduction in fucosylation, suggesting that HsPOFUT2 is highly selective for amino acids in the Xa position | Caenorhabditis elegans |
S15Q | site-directed mutagenesis, the mutant shows increased secretion, but a significant reduction in fucosylation, suggesting that HsPOFUT2 is highly selective for amino acids in the Xa position | Caenorhabditis elegans |
V16H | site-directed mutagenesis, the mutation affects both secretion and fucosylation | Caenorhabditis elegans |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | activates wild-type and mutant enzymes, but has a nonessential role in catalysis. Residue R290 replaces Mn2+ function by establishing electrostatic and hydrogen bond interactions with beta-phosphate | Caenorhabditis elegans |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Caenorhabditis elegans | Q8WR51 | gene profut2 | - |
Purification (Comment) | Organism |
---|---|
recombinant enzyme CePOFUT2 in complex with GDP and human TSR1 by gel filtration | Caenorhabditis elegans |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
GDP-beta-L-fucose + PCQNGGSCKDQL = PCQNGGS(O-beta-L-fucosyl)-CKDQL + GDP | catalytic mechanism of isozyme POFUT2 and its preference for threonine over serine residues | Caenorhabditis elegans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | enzyme POFUT2 fucosylates threonine preferentially over serine and relies on folded TSRs containing the minimal consensus sequence C-X-X-S/T-, substrate specificity, overview | Caenorhabditis elegans | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
POFUT2 | - |
Caenorhabditis elegans |
protein O-fucosyltransferase 2 | - |
Caenorhabditis elegans |
General Information | Comment | Organism |
---|---|---|
additional information | enzyme in complex with GDP and human thrombospondin type 1 repeat shows an inverting mechanism for fucose transfer assisted by a catalytic base and shows that nearly half of the thrombospondin type 1 repeat is embraced by CePOFUT2. A small number of direct interactions and a large network of water molecules maintain the complex, role of interstitial water in the complex interface, water-mediated interactions, overview | Caenorhabditis elegans |
physiological function | protein O-fucosyltransferase 2 is an essential enzyme that fucosylates serine and threonine residues of folded thrombospondin type 1 repeats | Caenorhabditis elegans |