Human Cytosolic Sulfotransferase Sub-Family 4A Member 1

Target ID:

SULT4A1

Aliases:

BR-STL-1BRSTL1DJ388M5.3hBR-STL-1MGC40032NSTSULTX3

Deposition Date:

Wednesday 13th April 2005

Families:

Transferases

Related Diseases:

MetabolismDrug metabolism and toxicologyCancer

Structure type:

apo

Download Coordinates:

1ZD1

Authors:

A.DONG,L.DOMBROVSKI,P.LOPPNAU,A.M.EDWARDS,C.H.ARROWSMITH,M.SUNDSTROM,A.BOCHKAREV,A.N.PLOTNIKOV,STRUCTURAL GENOMICSCONSORTIUM (SGC)

Site:

Toronto

ICM Datapack:

ICM Datapack

1ZD1

tabs

Structure Details

Human sulfotransferase SULT4A1 is a member of cytosolic sulfotransfeares protein family (SULTs),
which metabolize small molecules including various hormones and numerous xenobiotics.
It is also called 'brain sulfotransferase-like cDNA/protein' (BR-STL),
because the protein is expressed only in the brain.
So far, no substrate has been identified for SULT4A1 and the function of this enzyme remains unknown.

We have solved the crystal structure of SULT4A1 at 2.2Ã… resolution.
The crystal structure provides important information for rational search of SULT4A1 substrates
of endogenous and exogenous origin.

Materials and Methods

SULT4A1

PDB:1ZD1

Revision

Revision Type:created
Revised by:created
Revision Date:created
Entry Clone Accession:gi:7657633
Entry Clone Source:MGC
SGC Clone Accession:
Tag:N-terminal: His-tag with integrated thrombin protease site: MGSSHHHHHHSSGLVPR*GS
Host:E.coli BL21 (DE3) codon plus RIL (Stratagene)

Construct

Prelude:
Sequence:gsMAESEAETPSTPGEFESKYFEFHGVRLPPFCRGKMEEIANFPVRPSDVWIVTYPKSGTSLLQEVVYLVSQGADPDEIGLMNIDEQLPVLEYPQPGLDIIKELTSPRLIKSHLPYRFLPSDLHNGDSKVIYMARNPKDLVVSYYQFHRSLRTMSYRGTFQEFCRRFMNDKLGYGSWFEHVQEFWEHRMDSNVLFLKYEDMHRDLVTMVEQLARFLGVSCDKAQLEALTEHCHQLVDQCCNAEALPVGRGRVGLWKDIFTVSMNEKFDLVYKQKMGKCDLTFDFYL
Vector:p28a-LIC

Growth

Medium:
Antibiotics:
Procedure:SULT4A1 was expressed in E.coli BL21 (DE3) codon plus RIL in Terrific Broth (TB) in the presence of 50 µg/mL of kanamycin at 37 ºC to an OD600 of 0.8. Cells were then induced by isopropyl-1-thio-D-galactopyranoside (IPTG), final concentration 0.5 mM, and incubated overnight at 15 ºC.

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 (phosphate buffer saline (PBS), pH 7.4, 0.5 M NaCl, 5 mM imidazol, 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:SULT4A1 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 20% polyethylene glycol 4000, 0.2 M ammonium tartrate
NMR Spectroscopy:
Data Collection:
Data Processing: