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Literature summary extracted from
- Mapping of a substrate binding site in the protein disulfide isomerase-related chaperone Wind based on protein function and crystal structure (2004), J. Biol. Chem., 279, 39829-39837.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 5.3.4.1 | DNA sequence determination, expression of GFP-tagged wild-type enzyme and mutants Y53S and K84D in COS-7 cells, expression of C-terminally or N-terminally His6-tagged wild-type and mutant Wind in Escherichia coli strain XL 1-Blue and in Vero cells, the affinity for substrate Pipe is higher with the N-terminally His-tagged enzyme compared to the C-terminally tagged one | Drosophila melanogaster |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 5.3.4.1 | C24S | site-directed mutagenesis, unaltered Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | C27S | site-directed mutagenesis, unaltered Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | D29N | site-directed mutagenesis, unaltered Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | D31N | site-directed mutagenesis, unstable protein that still dimerizes but then aggregates, unaltered Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | D31N/R41S | site-directed mutagenesis, monomeric mutant, slightly decreased Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | D50A | site-directed mutagenesis, inactive mutant, normal dimerization | Drosophila melanogaster |
| 5.3.4.1 | D50N | site-directed mutagenesis, decreased Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | D50S | site-directed mutagenesis, altered Pipe targeting compared to the wild-type enzyme, normal dimerization | Drosophila melanogaster |
| 5.3.4.1 | D85N | site-directed mutagenesis, unaltered Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | E32K | site-directed mutagenesis, unaltered Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | E60A | site-directed mutagenesis, unaltered Pipe processing and targeting efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | E60Q | site-directed mutagenesis, decreased Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | E60Y | site-directed mutagenesis, unaltered Pipe processing and targeting efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | E88K | site-directed mutagenesis, unaltered Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | E88Q | site-directed mutagenesis, unaltered Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | E90R | site-directed mutagenesis, unaltered Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | E90R/D29N | site-directed mutagenesis, unaltered Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | E90R/E60A | site-directed mutagenesis, unaltered Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | G87S | site-directed mutagenesis, unaltered Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | H59Y | site-directed mutagenesis, unaltered Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | K58S | site-directed mutagenesis, unaltered Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | K84D | site-directed mutagenesis, decreased Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | K84N | site-directed mutagenesis, unaltered Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | K84S | site-directed mutagenesis, unaltered Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | L219S/E212Q | site-directed mutagenesis, decreased Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | L232K | site-directed mutagenesis, increased Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | P106D | site-directed mutagenesis, similar to the wild-type enzyme, altered conformation | Drosophila melanogaster |
| 5.3.4.1 | R215A | site-directed mutagenesis, unaltered Pipe processing efficiency but eliminated Pipe targeting compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | R218D | site-directed mutagenesis, increased Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | R41S | site-directed mutagenesis, sligtly reduced dimerization of the mutant and processing of Pipe | Drosophila melanogaster |
| 5.3.4.1 | T25K | site-directed mutagenesis, unaltered Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | T63K | site-directed mutagenesis, unaltered Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | V28D | site-directed mutagenesis, monomeric mutant, no dimerization, mutant aggregates | Drosophila melanogaster |
| 5.3.4.1 | V28Y | site-directed mutagenesis, monomeric mutant, no dimerization, mutant aggregates | Drosophila melanogaster |
| 5.3.4.1 | Y53S | site-directed mutagenesis, decreased Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | Y55K | site-directed mutagenesis, mutant shows increased affinity for substrate Pipe but decreased processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | Y55K/D31N/R41S | site-directed mutagenesis, slightly decreased Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | Y55S | site-directed mutagenesis, decreased Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | Y86F | site-directed mutagenesis, unaltered Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | Y86L | site-directed mutagenesis, highly decreased Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | Y86Q | site-directed mutagenesis, highly decreased Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
| 5.3.4.1 | Y86S | site-directed mutagenesis, unaltered Pipe processing efficiency compared to the wild-type enzyme | Drosophila melanogaster |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 5.3.4.1 | Drosophila melanogaster | - |
- |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 5.3.4.1 | recombinant His6-tagged wild-type and mutant Wind from Escherichia coli strain XL 1-Blue by nickel affinity chromatography | Drosophila melanogaster |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 5.3.4.1 | Pipe | processing and targeting of Pipe, Pipe is an essential Golgi transmembrane-O-sulfotransferase, protein disulfide isomerase-related chaperone Wind is required for processing and correct targeting of the substrate, mapping of multiple substrate binding sites in Pipe, one enzyme site in vicinity of an exposed cluster of tyrosine residues within the thioredoxin fold domain is essential for activity, a second enzyme site in the enzyme's D-domain is also necessary for processing activity, but competitive to the thioredoxin fold domain residue, overview | Drosophila melanogaster | ? | - |
? |  |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 5.3.4.1 | dimer | - |
Drosophila melanogaster |
| 5.3.4.1 | More | protein disulfide isomerase-related chaperone Wind contains a thioredoxin fold domain and a C-terminal D-domain unique in PDI-D proteins | Drosophila melanogaster |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 5.3.4.1 | PDI | - |
Drosophila melanogaster |
| 5.3.4.1 | protein disulfide isomerase | - |
Drosophila melanogaster |
| 5.3.4.1 | protein disulfide isomerase-related chaperone Wind | - |
Drosophila melanogaster |
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Release 2023.1 (January 2023)
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