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Literature summary for 3.2.2.22 extracted from

  • Guenhan, E.; Swe, M.; Palazoglu, M.; Voss, J.C.; Chalupa, L.M.
    Expression and purification of cysteine introduced recombinant saporin (2008), Protein Expr. Purif., 58, 203-209.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.2.2.22

Application

Application Comment Organism
pharmacology enables easy immunotoxin creation by coupling target antigen to artificial cysteine residue at non-enzyme-interfering C-terminus of toxin in a predictive way Saponaria officinalis

Cloned(Commentary)

Cloned (Comment) Organism
Escherichia coli XL-1 Blue (high transformation efficiency) and Bl21 (DE3) pLysS (high expression) with plasmid pET11d Saponaria officinalis

Protein Variants

Protein Variants Comment Organism
S255C intact stable enzyme, site far away from ribosome inactivating site residues, N-terminal mutations were unstable, low tendency to dimerization by disulfide-bonding, but high level of cross-linking with nonspecific mouse anti-human IgG antibody Saponaria officinalis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
30000
-
 SDS-PAGE, slightly higher than pure saporine-3 (253 amino acids) due to introduction of 7 additional amino acids yielding a 260 amino acid protein Saponaria officinalis

Organism

Organism UniProt Comment Textmining
Saponaria officinalis
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-
-

Purification (Commentary)

Purification (Comment) Organism
centrifugation, 0.22 microM filtered, dialyzed against 25 mM sodium chloride in phosphate buffer, pH 6.0, with 2 mM EDTA, application to HP CM FF Sepharose ion exchange column Saponaria officinalis

Source Tissue

Source Tissue Comment Organism Textmining
seed
-
 Saponaria officinalis
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 IC50 = 0.00000000084 mM for translation inhibition of mutated saporin-3 comparable to native saporin-3 isomer and commercial saporin as controls Saponaria officinalis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
mRNA + H2O in vitro translation inhibition assay in rabbit reticulocyte lysate, 30 degrees Celsius Saponaria officinalis ?
-
 ?

Subunits

Subunits Comment Organism
? x * 30000, SDS-PAGE Saponaria officinalis

Synonyms

Synonyms Comment Organism
saporin saporin-3 with cysteine 255 mutation at C-terminus Saponaria officinalis