| Cat. No. : |
SlyD-329E |
| Description : |
SlyD is a putative folding helper protein from the Escherichia coli cytosol, which consists of an N-terminal prolyl isomerase domain of the FKBP type and a presumably unstructured C-terminal tail. It is involved in the biosynthesis of the metal cluster in the [NiFe]-hydrogenase enzymes, and exhibits several activities including that of a peptidyl-prolyl isomerase. |
| Source : |
E.coli. |
| Formulation : |
Liquid. In 20 mM Tris-HCl buffer (pH 7.5). |
| Molecular Weight : |
21 kDa (196 aa). |
| Purity : |
> 95% by SDS – PAGE. |
| Concentration : |
1 mg/ml (determined by Bradford assay). |
| Sequence : |
MKVAKDLVVS LAYQVRTEDG VLVDESPVSA PLDYLHGHGS LISGLETALE GHEVGDKFDV AVGANDAYGQ YDENLVQRVP KDVFMGVDEL QVGMRFLAET DQGPVPVEIT AVEDDHVVVD GNHMLAGQNL KFNVEVVAIR EATEEELAHG HVHGAHDHHH DHDHDGCCGG HGHDHGHEHG GEGCCGGKGN GGCGCH |
| Biological activity : |
Specific activity is > 220 nmoles/min/mg, and is defined as the amount of enzyme that cleaves 1umole of suc-AAFP-pNA per minute at 25°C in Tris-Hcl pH8.0 using chymotrypsin. |
| Activity Assay : |
1. Prepare 170ul assay buffer into a suitable container and pre-chill on ice before use: The final concentrations are 200 mM Tris-Hcl, pH 8.0, and 20nM chymotrypsin.2. Add 10ul of recombinant SlyD protein with 1ug in assay buffer. 3. Mix by inversion and equilibrate to 1°C and monitor the A405nm until the value is constant using a spectrophotometer.4. Add 20ul pre-chilled 5mM suc-AAFP-pNA. (Substrate was dissolved in TFE that contained 460mM LiCl to a concentration of 3 mM)5. Record the increase in A405 nm for 30 minutes at 25°C. |
| Storage : |
Can be stored at +4°C short term (1-2 weeks). For long term storage, aliquot and store at -20°C or -70°C. Avoid repeated freezing and thawing cycles. |
