PMID- 9325148
OWN - NLM
STAT- MEDLINE
DCOM- 19971121
LR  - 20061115
IS  - 1046-5928 (Print)
IS  - 1046-5928 (Linking)
VI  - 11
IP  - 1
DP  - 1997 Oct
TI  - High level expression and characterization of recombinant human hippocampus 
      phenol sulfotransferase: a novel phenol-sulfating form of phenol 
      sulfotransferase.
PG  - 125-34
AB  - Phenol sulfotransferases (PSTs) represent a family of sulfotransferase enzymes 
      that modify the biologic activities and excretion of phenolic compounds and 
      monoamines. A novel human hippocampal PST (H-PST) cDNA with homology to phenol 
      (P) and monoamine (M) forms of PST was previously isolated from brain. To compare 
      the biochemical properties of H-PST with that of phenol (P-PST) and monoamine 
      (M-PST) sulfotransferases, high level expression of recombinant H-PST was 
      achieved in this study with the pET3c vector in BL21(DE3) Escherichia coli cells. 
      Expression was demonstrated by isopropyl beta-D-thiogalactopyranoside induction 
      of 34-kDa H-PST that represented 5-10% of total E. coli proteins. Purification by 
      ion-exchange chromatography on DEAE-Sepharose yielded more than 2 mg of H-PST. 
      Characterization showed that H-PST exists as a homodimer of 60-65 kDa by gel 
      filtration chromatography. H-PST prefers p-nitrophenol as substrate and does not 
      sulfate dopamine or neuropeptide substrates. Kinetic studies showed that H-PST 
      possessed K(m(app)) and Vmax(app) values of 3 microM p-nitrophenol and 160 
      nmol/min/mg, respectively. H-PST was sensitive to inhibition by DCNP 
      (2,6-dichloro-4-nitrophenol). H-PST is thermolabile since its activity was 
      reduced upon preincubation at 37 degrees C. These results indicate that H-PST 
      shows similarities and differences compared to P-PST and M-PST sulfotransferases. 
      P-PST prefers p-nitrophenol as substrate, is sensitive to inhibition by DCNP, and 
      is thermostable; in contrast, M-PST prefers monoamines as substrate, is not 
      sensitive to DCNP, and is thermolabile. The distinct profile of biochemical 
      properties of H-PST, and its primary sequence homology to P-PST and M-PST, 
      suggests that H-PST represents a novel allelic variant of human phenol 
      sulfotransferases. Importantly, this study demonstrates that high level 
      expression of H-PST allows determination of distinguishing characteristics of 
      variant forms of PSTs.
FAU - Hwang, S R
AU  - Hwang SR
AD  - Department of Medicine, University of California, San Diego 92093, USA.
FAU - Palkovits, M
AU  - Palkovits M
FAU - Hook, V Y
AU  - Hook VY
LA  - eng
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Protein Expr Purif
JT  - Protein expression and purification
JID - 9101496
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Isoenzymes)
RN  - 0 (Nitrophenols)
RN  - 0 (Recombinant Proteins)
RN  - 618-80-4 (2,6-dichloro-4-nitrophenol)
RN  - EC 2.8.2.1 (Arylsulfotransferase)
SB  - IM
MH  - Alleles
MH  - Amino Acid Sequence
MH  - Arylsulfotransferase/antagonists & inhibitors/*biosynthesis/chemistry/genetics
MH  - Blood Platelets/enzymology
MH  - Enzyme Inhibitors/pharmacology
MH  - Escherichia coli
MH  - Hippocampus/*enzymology
MH  - Humans
MH  - Isoenzymes/*biosynthesis/chemistry
MH  - Kinetics
MH  - Liver/enzymology
MH  - Molecular Sequence Data
MH  - Nitrophenols/pharmacology
MH  - Recombinant Proteins
MH  - Sequence Alignment
EDAT- 1997/11/05 00:00
MHDA- 1997/11/05 00:01
CRDT- 1997/11/05 00:00
PHST- 1997/11/05 00:00 [pubmed]
PHST- 1997/11/05 00:01 [medline]
PHST- 1997/11/05 00:00 [entrez]
AID - S1046-5928(97)90782-8 [pii]
AID - 10.1006/prep.1997.0782 [doi]
PST - ppublish
SO  - Protein Expr Purif. 1997 Oct;11(1):125-34. doi: 10.1006/prep.1997.0782.