SULT1C2
PDB:2AD1
Revision
Revision Type:created
Revised by:created
Revision Date:created
Entry Clone Accession:gi: 28830308
Entry Clone Source:MGC
SGC Clone Accession:
Tag:N-terminal: His-tag with integrated thrombin protease site: MGSSHHHHHHSSGLVPRGS
Host:E.coli BL21 (DE3) codon plus RIL (Stratagen).
Construct
Prelude:
Sequence:
Vector:p28a-LIC
Growth
Medium:
Antibiotics:
Procedure:
Purification
Procedure
Extraction
Procedure
Cells were harvested by centrifugation at 6,000 rpm. The cell pellets were frozen in liquid nitrogen and stored at -80°C. For the purification the cell paste was thawed and resuspended in lysis buffer (50 mM HEPES-NaOH, pH 7.4, 0.5 M NaCl, 5 mM imidazol, 2 mM ß-mercaptoethanol, 5% glycerol) with protease inhibitor (0.1mM phenylmethyl sulfonyl fluoride, PMSF). The cells were lysed by passing through Microfluidizer (Microfluidics Corp.) at 20,000 psi.
Concentration:
Ligand
MassSpec:
Crystallization:Purified SULT1C2 was crystallized using the hanging drop method at 20°C by mixing 2 µl of the protein solution with 2 µl of the reservoir solution containing 14-20% polyethylene glycol 3350, 0.2 M Li-Citrate, 0.1 M Na-Citrate, pH 4.6.
NMR Spectroscopy:
Data Collection:
Data Processing:
