slyD (1-196) Escherichia coli Protein

Specifications

Product Data
Species	Escherichia coli
Expression Host	E. coli
Expression cDNA Clone or AA Sequence	Protein Sequence (showhide) MKVAKDLVVS LAYQVRTEDG VLVDESPVSA PLDYLHGHGS LISGLETALE GHEVGDKFDV AVGANDAYGQ YDENLVQRVP KDVFMGVDEL QVGMRFLAET DQGPVPVEIT AVEDDHVVVD GNHMLAGQNL KFNVEVVAIR EATEEELAHG HVHGAHDHHH DHDHDGCCGG HGHDHGHEHG GEGCCGGKGN GGCGCH
Predicted MW	21 kDa
Concentration	lot specific
Purity	>95% by SDS PAGE
Buffer	Presentation State: Purified State: Liquid purified protein Buffer System: 20 mM Tris pH 7.5
Preparation	Liquid purified protein
Applications	Protocol: Activity Assay 1. Prepare 170 ul 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 10 ul of recombinant SlyD protein with 1 ug 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 20 ul 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.
Protein Description	Recombinant E.coli slyD was expressed in E.coli and purified by conventional chromatography.
Storage	Store undiluted at 2-8°C for up to two weeks or (in aliquots) at -20°C or -70°C for longer. Avoid repeated freezing and thawing.
Stability	Shelf life: one year from despatch.
Bioactivity	Specific: > 220 nmoles/min/mg, defined as the amount of enzyme that cleaves 1 umole of suc-AAPF-pNA per minute at 25°C in Tris-HCl pH 8.0 using chymotrypsin.
Reference Data
Summary	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.

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