PMID- 9497349
OWN - NLM
STAT- MEDLINE
DCOM- 19980407
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 273
IP  - 11
DP  - 1998 Mar 13
TI  - Localization and functional analysis of the substrate specificity/catalytic 
      domains of human M-form and P-form phenol sulfotransferases.
PG  - 6242-7
AB  - Human monoamine (M)-form and simple phenol (P)-form phenol sulfotransferases 
      (PSTs), which are greater than 93% identical in their primary sequences, were 
      used as models for investigating the structural determinants responsible for 
      their distinct substrate specificity and other enzymatic properties. A series of 
      chimeric PSTs were constructed by reciprocal exchanges of DNA segments between 
      cDNAs encoding M-form and P-form PSTs. Functional characterization of the 
      recombinant wild-type M-form, P-form, and chimeric PSTs expressed in Escherichia 
      coli and purified to homogeneity revealed that internal domain-spanning amino 
      acid residues 84-148 contain the structural determinants for the substrate 
      specificity of either M-form or P-form PST. Data on the kinetic constants (Km, 
      Vmax, and Vmax/Km) further showed the differential roles of the two highly 
      variable regions (Region I spanning amino acid residues 84-89 and Region II 
      spanning amino acid residues 143-148) in substrate binding, catalysis, and 
      sensitivity to the inhibition by 2,6-dichloro-4-nitrophenol. In contrast to the 
      differential sulfotransferase activities of M-form and P-form PSTs toward 
      dopamine and p-nitrophenol, the Dopa/tyrosine sulfotransferase activities were 
      found to be restricted to M-form, but not P-form, PST. Furthermore, the variable 
      Region II of M-form PST appeared to play a predominant role in determining the 
      Dopa/tyrosine sulfotransferase activities of chimeric PSTs. Kinetic studies 
      indicated the role of manganese ions in dramatically enhancing the binding of 
      D-p-tyrosine to wild-type M-form PST. Taken together, these results pinpoint 
      unequivocally the sequence encompassing amino acid residues 84-148 to be the 
      substrate specificity/catalytic domain of both M-form and P-form PSTs and 
      indicate the importance of the variable Regions I and II in determining their 
      distinct enzymatic properties.
FAU - Sakakibara, Y
AU  - Sakakibara Y
AD  - Department of Biochemistry, University of Texas Health Center, Tyler, Texas 
      75710, USA.
FAU - Takami, Y
AU  - Takami Y
FAU - Nakayama, T
AU  - Nakayama T
FAU - Suiko, M
AU  - Suiko M
FAU - Liu, M C
AU  - Liu MC
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Nitrophenols)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 42Z2K6ZL8P (Manganese)
RN  - 618-80-4 (2,6-dichloro-4-nitrophenol)
RN  - EC 2.8.2.1 (Arylsulfotransferase)
RN  - VTD58H1Z2X (Dopamine)
RN  - Y92ZL45L4R (4-nitrophenol)
SB  - IM
MH  - Amino Acid Sequence
MH  - Arylsulfotransferase/drug effects/genetics/isolation & purification/*metabolism
MH  - Binding Sites
MH  - Dopamine/metabolism
MH  - Escherichia coli/genetics
MH  - Humans
MH  - Kinetics
MH  - Manganese/pharmacology
MH  - Molecular Sequence Data
MH  - Nitrophenols/metabolism/pharmacology
MH  - Recombinant Fusion Proteins/metabolism
MH  - Sequence Homology, Amino Acid
MH  - Substrate Specificity
EDAT- 1998/04/16 00:00
MHDA- 1998/04/16 00:01
CRDT- 1998/04/16 00:00
PHST- 1998/04/16 00:00 [pubmed]
PHST- 1998/04/16 00:01 [medline]
PHST- 1998/04/16 00:00 [entrez]
AID - S0021-9258(18)67737-3 [pii]
AID - 10.1074/jbc.273.11.6242 [doi]
PST - ppublish
SO  - J Biol Chem. 1998 Mar 13;273(11):6242-7. doi: 10.1074/jbc.273.11.6242.