THE JOURNALOF BIOLOGICAL

CHEMISTRY
Val. 256, No. 21, Issue of November 10, pp. 11123-11127, 1981
Printed in U.S.A.

On the Mechanism of ArylSulfotransferase*

(Received for publication, March 25, 1981, and in revised form, June 18, 1981)

Michael W. Duffel and William B.Jakoby

From theSection on Enzymes and Cellular Biochemistry, National Institute of Arthritis, Metabolism, a n d Digestive
Diseases, National Institutes of Health, Bethesda, Maryland 20205

Aryl sulfotransferase IV (EC 2.8.2.1), purified to ho- requirement for P A P the reaction (Equation 2) was experi-
mogeneity from male rat liver, catalyzes the sulfation mentally irreversible. Although this “exchange” reaction has
of a varietyof substituted phenols, including catechol- been applied as an assay for sulfotransferase from several
amines, tyrosine esters, and peptides containing NH2- tissues, all aryl sulfotransferases do not catalyze it (2).
terminal tyrosine residues. An investigation of the Although incomplete kinetic studies have been conducted
mechanismoftheenzyme was carriedout using 2- with partially purified sulfotransferases ( 7 , 8 ) ,the availability
chloro-4-nitrophenolas a model substrate. of an homogeneous enzyme preparation ( 6 ) has allowed us to
Kinetic, inhibition,and bindingstudies with aryl sul- investigate more thoroughly the kinetic and chemical mech-
fotransferase IV are all consistent with a random rapid anism of the PAPS-dependent sulfate transfer reaction. We
equilibrium BiBi kinetic mechanism with two dead end have also examined the PAP-requiring exchange reaction, as
product inhibitor complexes. Studies of the chemical well as a nucleotide-independent hydrolytic activity, that are
mechanisms of the enzyme-catalyzed reaction demon- catalyzed by the same enzyme.
strate that electron-withdrawing substituents decrease
the maximalvelocity of phenol sulfation.The maximal MATERIALS AND METHODS
velocity of the reaction correlates with Hammett 0,- Substrates and Products-CNP, pentachlorophenol, and all 4-sub-
constants ( p = -0.25). Evidence is presented for the stituted phenols were purchased from Aldrich. I-Naphthol, 2-naph-
mechanism by which adenosine 3’,5’-bisphosphate and thol, 1-naphthyl sulfate, 4-nitrophenyl sulfate, and PAP were ob-
aryl sulfotransferase catalyze the transfer of sulfate tained from Sigma. All phenols and phenyl sulfates were either
from 2-chloro-4-nitrophenyl sulfate to other phenols. distilled or recrystallized prior to use. PAPS was prepared syntheti-
cally (4); [%3]PAPS was purchased from New England Nuclear and
purified as described (10).
CNPS was synthesized by sulfation of CNP with chlorosulfonic
The formation of sulfate esters in mammalian tissues is a acid in a modification of a published procedure (11).Sulfation was
major pathway of metabolism for substrates bearing a hy- carried out in tetrahydrofuran and N,N-dimethylaniline. After isola-
droxyl functional group (1-3), and serves as ameans for tion as a potassium salt, the product was dissolved in 0.25 M sodium
preparing lipophilic xenobiotics for excretion (2, 3 ) . While acetate, pH 3.5, a t 4 “C and the solution rapidly extracted four times
with ethyl ether. The aqueous layer, a t 4 “C, was quickly adjusted to
many compounds have been found to serve as acceptors in pH 7 with sodium hydroxide and stored at -20 “C toprevent sulfate
the sulfotransferase reaction inwhich adenosine 3”phosphate ester hydrolysis. This procedure yielded CNPS with less than 0.03%
5’-phosphosulfate acts as the sulfate donor (Equation l), iden- contamination by CNP.
tification of the enzymes responsible required separation of Aryl Sulfotransferase ZV-Aryl sulfotransferase IV was purified
the several catalytic species (3). This has been accomplished from the livers of male Sprague-Dawley rats aspreviously described
for enzymes with specificity for bile acids, for phenolic ste- (6). After chromatography on hydroxylapatite, aryl sulfotransferase
IV was completely separated from aryl sulfotransferase 111 by prepar-
roids, for primary and secondary alcohols including hydroxy- ative isoelectric focusing in a sucrose gradient. A linear pH gradient
steroids, and for phenols ( 3 ) .At least two types of aryl sulfo- from 3.5 to 10 (LKB Ampholines) wasused in a 110-ml column
transferase that differ somewhat in substrate specificity have (LKB); electrofocusing was carried out for 16 h at 15 watts (constant
been identified for phenols (3-6) and have been purified to power). The peak activity for sulfotransferase IV was eluted in frac-
homogeneity from rat liver (5, 6 ) . The enzyme used for the tions which were at pH 5.6 to 5.8. This was well separated from the
studies described here, arylsulfotransferase IV (3-6),catalyzes peak aryl sulfotransferase I11 activity at pH 6.4. The separation of
the reversible sulfation of a large variety of substituted phe- sulfotransferases IV and 111 by these means was essential for kinetic
studies since the previously described method required an affinity
nols, and is active with organic hydroxamic acids, catechol- step in which PAPS was used (6); exposure of the enzyme to this
amines, and tyrosine esters. substrate would have disqualified the enzyme preparation for the
ROH + PAPS’ + ROS03- + H’ + PAP (1)
kinetic studies conducted here. The resulting aryl sulfotransferase IV
was homogeneous upon sodium dodecyl sulfate gel electrophoresis
4-Nitrophenyl sulfate and had the same specific activity (630 nmol min” mg of protein”)
+ phenol zp4-nitrophenol + phenyl sulfate (2) at pH 5.5 with 2-naphtho1, as did preparations of the enzyme purified
by affinity chromatography. After isoelectric focusing the enzyme was
In addition to the fist reaction (Equation I), Gregory and stored at 4 “C in the presence of 5 mM 2-mercaptoethanol and 3mM
Lipmann (9) described the transfer of sulfate from 4-nitro- sodium azide.
phenyl sulfate to phenol in the absence of PAPS, but with a Initial Rate Kinetics-Initial rate studies were carried out by
measuring the change in absorbance a t 400 nm due to CNP with a
* The costs of publication of this article were defrayed in part by Cary 219 spectrophotometer equipped with a temperature-controlled
the payment of page charges. This article must therefore be hereby cell holder maintained at 25 “C. Assay mixtures contained 200 mM
marked “advertisement” in accordance with 18 U.S.C. Section 1734 potassium phosphate (pH 7.0), 5 mM 2-mercaptoethanol, and varying
solely to indicate this fact. concentrations of PAPS, PAP, CNP, CNPS, and arylsulfotransferase
’ The abbreviations used are: PAPS, adenosine 3-phosphate 5’- IV in a total volume of 1.0 ml. Concentrations were calculated from
phosphosulfate; PAP, adenosine 3’,5’-bisphosphate;CNP, 2-chloro-4- the molar absorptivity, 10.1 X IO? liters/mol” cm”for CNP. All
nitrophenol; CNPS, 2-chloro-4-nitrophenyl sulfate. velocities are expressed as micromoles of CNP produced or consumed

11123

This is an Open Access article under the CC BY license.

 -F”!
11124 Mechanism
SulfotransferaseAryl
per min. The molar ratio of limiting substrate to enzyme was greater 1 K N P X 10’ IvM1.l liPAPS x lo2 IvMI ’
than 100 to 1, except in those cases where substrate concentration 0 20 40
was between 3 and 6 p ~ i.e. , 301 or greater. Kinetic constants were

/i
obtained from slope and interceptreplots of the initial rate data. The
standard errors associated with these constants were less than 10%.
Protein concentrations were measured by a modified Lowryprocedure
loo!A
(12) in which bovine serum albumin served as standard.
Binding of PAPS to Aryl Sulfotransferase IV-In the absence of
acceptor phenols, binding of PAPS to theenzyme was measured with
a Farrand Mark I spectrofluorimeter equipped with a temperature-
controlled cell compartment maintained at 25 “C. Cuvettes containing
140 p l of 0.14 M potassium phosphate (pH 7.0), 3.5 mM 2-mercapto-
ethanol, and 2.0 p~ aryl sulfotransferase IV were allowed to equili-
brate for 5 min. A maximum of 16 pl of 2 mM PAPS was added in 2-,
4-, or 8-pl aliquots, and the solution was stirred with a Teflon rod
after each addition. The decrease in intrinsic fluorescence of the
protein was measured at 345 nm (excitation at 285 nm) after each
addition of PAPS. Absorption of incident or fluorescent light by
PAPS under these conditions did not significantly change the fluo-
rescence of the protein; the datawere corrected for the small dilution
occurring when PAPS was added.
Sulfation of 4-substituted Phenols-Maximal velocities for sulfa-
0‘
tion of 4-substituted phenols were obtained by varying the concentra- 0 155 10 0 10 M
tion of the phenol and PAPS in a constant ratio (13). PAPS concen-
trations were 9.3 PM, 18.5 p ~ 37.0
, p ~ and, 74.0 p ~ The
. concentration
l/CNPSx 10’ IpMI.’ l/PAP x 10’ ipM) ’
ratio used for substituted phenol to PAPS was 70 for p-cresol, 80 for FIG. 1. Initial rate data for aryl sulfotransferase IV.Reaction
4-methoxyphenol, 50 for phenol, 20 for 4-chlorophenol, 5 for methyl- conditions are described under “Materials and Methods.” In A, the
4-hydroxybenzoate, 5 for 4-hydroxyacetophenone, and 1 for 4-nitro- concentrations of PAPS (top to bottom) were 5.4 pM, 10.8 pM, 21.6
phenol. Assays were conducted with a [%]PAPS chromatographic PM, 43.2 p ~ and
, 86.4 p ~ In. B , the concentrations of CNP were 3.3
procedure (10) using10 min of incubation at 25 “C and pH 7.0. p ~ 6.6
, pM, 13.4 p ~ 26.9 , 53.8 PM. For A and B , final enzyme
, p ~ and
Nonlinear regression analysis was performed with an interactive concentration was 1.2 X lo-’ M. In C, the concentrations of PAP were
curve-fitting program (14) on a Decsystem-10 computer. The maximal 4.9 PM, 9.8 p ~ 19.7
, p ~ 39.4
, p ~ and, 78.7 p ~ In. D,the concentrations
velocities were correlated with Hammett 0,- constants that hadbeen of CNPS were 8.6 p ~ 17.2 , p ~ 34.4
, p ~ 68.8 . C
, phf3 and 128 p ~ For
determined on the basis of phenol ionization (15). A least squares fit and D, final enzyme concentration was 6 X lo-’ M. The theoretical
of the data toa straight line yielded a value for p. lines were calculated from the appropriate sequential rate equation
(Equation 4).
RESULTS

Substrate Kinetics-The kinetic mechanism for the reac- A 10

tion of PAPS and 2-chloro-4-nitrophenolto form PAP and 2-
chloro-4-nitrophenyl sulfate (Equation 31, is judged to be
sequential on the basis of the data for the forward and reverse
CNP + PAPS $ CNPS + PAP (3)
directions inFig. 1. The velocity ( u ) in either direction is
predicted by the general rate equation (Equation 4) in which
A and B aresubstrates, and K,,, K,, and K b are kinetic
constants (16) that may be calculated from slope and intercept
replots of the initial rate data. Since the replots were linear,
the kinetic constants obtained from them (Table I) were used
to generate the theoretical lines in Fig. 1.
0
0 0.05 0.1 0 0.1 0.2
1 /CNPS (~“’1 1/PAP (FM”)
TABLEI FIG. 2. Product inhibitiondata for aryl sulfotransferaseIV.
Kinetic constants for aryl sulfotransferase IV calculatedfor a Reaction conditions were the same as for initial rate kinetics. Final
rapid equilibrium random mechanism enzyme concentration was 6 X lo-@M. In I, PAP concentration was
19.7 p ~CNP . concentrations were a, no CNP; b, 3.6 p ~c,;7.2 p ~ and
;
E-PAPS-CNPS d, 14.4 PM. In ZI, CNPS concentration was 34.4 p ~ CNP . concentra-
; h, 14.4 p ~ In. 111, PAP
tions were e, no CNP; f, 3.6 p ~ g,;7.2 p ~ and
E-PAPS
K
concentration was 19.7 p ~ PAPS
. concentrations were i, no P A P S j ,
105 p ~k ,; 210 p ~ and
; 1,420 p ~ In. IV, CNPS concentration was 34.4
p ~ PAPS
. concentrations were m, no PAPS; n, 42 pM; 0,105 pM; and
E-PAPS-CNP E 5E E-PAP-CNPS p , 210 p ~ The. lines were calculated by the method of least squares.

Kinetics of Inhibition-Although the data of Fig. 1 are

formally inconsistent with a ping-pong mechanism, inhibitor
studies are necessary for further delimiting the number of
4 +
possible kinetic pathways. Data for product inhibition by CNP
Vm = 2 8 n r n 0 I m i n - ~ r n g - . ‘ Vm = 582nmolmin“mg” and PAPS are shown in Fig. 2; similar data are available for
inhibition of the forward reaction (Equation 3) by CNPS and
PAP, but are not presented. Table I1 presents a summary of
the patterns of inhibition when aryl sulfotransferase IV is
exposed to a dead end inhibitor, pentachlorophenol (17), and
 Aryl
Mechanism
Sulfotransferase 11125
TABLE I1 acceptor phenol, and under conditions in which the concen-
Inhibition of aryl sulfotransferase ZV trations of sulfotransferase and PAP were similar, there was
Varied substrate“ an essentially stoichiometric transfer of sulfate from CNPS to
Inhibitor PAP asmeasured by the increased absorbance accompanying
PAPS
CNP PAP CNPS
the formation of CNP. PAP could not be replaced by PAPS.
PAP Competi- Mixedb The reaction is demonstrated in phase I of Fig. 4, where the
tive
CNPS Mixed Competi-
tive
PAPS Competi- Mixed
tive
CNP Mixed Competi-
tive
Pentachloro- Mixed Competi-
phenol tive
” In all cases the substrate held constant was not saturating.
‘Both the slope and the intercept of the double reciprocal plot
change with changing inhibitor concentration.

to anyof the possible products of the reaction. The patternof

product inhibition is consistent with the rapid equilibrium
random mechanism with two deadend complexes that is
illustrated in the scheme presented inTable I. The patternof
product inhibition,but not thescheme, is also consistent with
each of two orderedTheorell-Chance mechanisms: in one 1/PAPS ( m M ) ”
case, PAPS would bind to theenzyme frst; in the other, CNP FIG. 3. Bindingof PAPS to aryl sulfotransferase IV. The
would bind first. decrease in fluorescence measured at 345 nm (AF) is expressed in
arbitrary units. The dissociation constant derived for PAPS binding
As indicated in Table 11, inhibition by pentachlorophenol
is 45 f 5 p ~ .
was competitive with CNP and could be defined as dead end
inhibition since pentachlorophenyl sulfate could not be de-
tected as a productby a sensitive chromatographic assaywith
[”SIPAPS (10). Since pentachlorophenol exhibited mixed 2.0 1 / “I
inhibition with respect to PAPS, the data are inconsistent
with an ordered mechanism in which PAPS is the frst sub-
strate to add toenzyme. A dissociation constant of 0.2 ~ L Mfor
enzyme and pentachlorophenol was estimated from the inhi-
bition data.
Binding of PAPS to Enzyme-Advantage was taken of the
intrinsic fluorescence of tryptophan in aryl sulfotransferase
IV in order to titrate changes in fluorescence as a function of a.PAPS+ROH ;
+ ROSJ;+H*+PAP

binding of PAPS to thetransferase.‘ The decrease in fluores- CNPS+PAP Y CNP+PAPS

cence upon addition of PAPS, in the absence of a phenol 0 15 5 10

substrate (Fig. 3), allowed the estimation of a PAPS-enzyme Time (min)
dissociation constant of 45 f 5 p ~ This
. dissociation constant
compared favorably with that for the PAPS-enzyme dissocia- FIG. 4. Stoichiometry for sulfation of PAP. Assay mixtures
contained 0.20 M potassium phosphate (pH 7.0), 5 mM Z-mercaptoeth-
tion, the latter obtained by replots of initial rate data (Kia in anol, 0.4 p~ PAP, 0.19 mM CNPS, and either 14.4 pg (- - -) or 7.2 pg
Table I), and is consistent with the assumption that this (-) of aryl sulfotransferase IV in a total volume of 1.0 ml. Forma-
binding is at a productive site. Sincebinding of enzyme with tion of CNP was measured by the change in absorbance at 400 nm.
either PAPS or CNP can take place in the absence of the ..
Extrapolation (. . ) of the PAP-independent rate (phase Z Z ) yields
respective second substrate, only arandom mechanism is the amount of CNP formed due to conversion of PAP to PAPS. Phase
consistent with the data. ZZZ is the result of adding 5 p~ 2-naphthol to the reaction mixture.
Hydrolysis of Aryl Sulfate-In the absence of both PAP
and PAPS, incubation of CNPS with the sulfotransferase led I I I I I 1 1 I I I 1 I

to hydrolysis of the substrate (Equation5),

CNPS + H20 CNP + H2S04 (5)

which could be readily measured as increased absorbance at

400 nm due to theformation of free phenol. By this means, an
apparent K , of 260 p~ was obtained for CNPS asa sulfatase
substrate at a maximal velocity that is 1.2% of that for the
reverse of Equation 1. The sulfatase activity is strictly de-
pendent on sulfotransferase, and is inhibited by pentachloro- 0 1 2 3 4 0 2 4 6 8 1 0
phenol. l/PhenolImM)”
1/ 2-Naphthol x 10 (rM)
The “Exchange” Reaction-The transfer of sulfate from FIG. 5. Transfer of sulfate from CNPS to phenol or 2-naph-
CNPS to an acceptor phenol required the presence of catalytic thol. Assay mixtures contained 0.2 M potassium phosphate (pH 7.0),
amounts of PAP, suggesting that Reaction 2 takes place in 5 mM 2-mercaptoethanol, 0.1 ~ L MPAP, 0.19 mM CNPS, 7.2 pg of aryl
two stages (see the equations in Fig. 4). In the absence of an sulfotransferase IV, and the indicated concentrations of phenol or 2-
naphthol. PAP was added last, and, after the initial stoichiometric
Due to fluorescence quenching by DNP, a similar binding study formation of PAPS, the PAP-independent CNPS sulfatase rate was
could not be carried out with this substrate. subtracted from the steady state rate.
11126 Aryl Sulfotransferase Mechanism
random Bi Bi mechanism that allows two dead end product
-0.2 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 inhibitor complexes (the scheme shown in Table I), or with
‘
1 an ordered Theorell-Chance mechanism. On the basis of 1)
binding studies with PAPS, 2) the binding of CNPS inde-
pendent of nucleotide, i.e. the sulfatase activity, and 3) the
8
kinetics of pentachlorophenol inhibition, the mechanism ap-
pears to correspond to thatpresented in Table I.
On the basis of the four Haldane equations in effect for a
1.6
rapid equilibrium random mechanism (Equation 6), kinetic
constants (Table I)may be used to approximate the equilib-
rium constant for the enzyme-catalyzed sulfation of CNP
(Equation 3). From the four equations (Equation 6), K,, was
found to be 0.045 f 0.009. Thus, at pH 7.0, Equation3
represents an endergonic reaction with a change in free energy
of +1.8 kcal per mol.

R
FIG. 6 . Effect of substituents on phenol sulfation. Hammett The two reactions controlling the PAP-dependent transfer
plot based on Vmaxdata for sulfation of p-substituted phenols. of sulfate from CNPS to anacceptor phenol are presented in
Fig. 4 (phase 111).Since PAP is a product inhibitor of the
observed stoichiometry between CNP and PAPis somewhat forward reaction, transfer from CNPS takes place only at a
greater than expected, 125%f 15%for the range of 0.05 to 0.4 very low concentration of PAP. Similarly, high concentrations
p~ PAP. In arriving at the stoichiometry, correction was of phenols with very low K, values, e.g. 1- and 2-naphthol,
required for the nucleotide-independent sulfatase activity of inhibit the formation of PAPS and, therefore, the transfer
the enzyme (Fig. 4, phase I I ) that also led to CNPformation. reaction. This inhibition at 5 to 10 p~ naphthol explains the
Upon addition of a sulfate acceptor, 2-naphthol in this case inability of previous investigators (7,9) todemonstrate trans-
(Fig. 4, phase I I I ) , the rate of formation of CNP was greatly fer from a phenyl sulfate to a naphthol, despite achieving
increased concomitant with the production of 2-naphthyl sul- sulfate transfer to phenol itself (9). In contrast to the naph-
fate. thols, phenol has a much higher K,, is not inhibitory, and
The PAP-mediatedtransfer of sulfate from CNPS to a serves as an acceptor at normal substrate concentrations.
second phenol has been demonstrated with aryl sulfotransfer- Thus, failure to transfer is due to inhibition phenomena rather
ase IV for 2-naphthol, 1-naphthol, and phenol. However, 2- than being based on the thermodynamic grounds of a “sulfate
naphthol was apotentproductinhibitor for the phase I potential” (9).
reaction (Fig. 4 ) , such that the transfer was appreciably de- The correlation of Vmaxwith up- values of phenols with p-
creased at concentrations greaterthan 10 p~ naphthol (Fig. 5, substituents indicates that resonance delocalization of nega-
right); transfer to 1-naphthol was similarly inhibited at con- tive charge on the phenol oxygen influences the rate-deter-
centrations above 5 PM. Phenol, however, is a very poor mining step of the reaction. The observed negative value of
inhibitor of the reverse reaction anddid not act asa substrate p, -0.25, suggests that electron-withdrawing substituents in-
inhibitor; the concentration of phenol necessary for half-max- crease the energy of the transition state of the reaction,
imal sulfate transfer under these conditions was 0.5 mM (Fig. thereby decreasing the V,,,,,. If, as for manyendothermic
5, left). High pressure liquid chromatography of reactants and reactions (18), the transition state resembles the products
products in the sulfate transfer reaction confiied that aryl more than the reactants, the presence of the SOs- group next
sulfotransferase IV catalyzed the formation of 2-naphthyl to the phenolic oxygenwould decrease the importance of
sulfate from CNPS, PAP, and 2-naphthol in the absence of resonance interactions with p-substituents. A small value of
exogenous PAPS. p for the reaction would result. Furthermore,the small p value
Effect of Substitution on Sulfation of Phenols-A group of suggests that the effect of electron-withdrawing substituents
p-substituted phenols have been chosen for an evaluation of is not directly on phenol ionization; for the latter, a much
the effect of electrophilicity on the enzymatic reaction rate. larger p would be expected. Instead, the effect of electron-
As evaluated kinetically by nonlinear regression analysis, withdrawing substituents is more likely to be on the transition
lower maximal velocities for enzyme-catalyzed reactions were state of the reaction, in agreement with studies (19) on the
the result of increasing the electron-withdrawing capacity of mechanism of the acid-catalyzed hydrolysis of p-nitrophenyl-
p-substituents. Quantitatively, theHammett up- constants sulfate; the small value of p (+0.5) observed for acid hydrolysis
correlated reasonably with the log of the maximum velocity was explained on the basis of the strongly electron-withdraw-
( p = -0.25; r = 0.89) (Fig. 6). The data did not correlate with ing SOa- group immediately adjacent to the reaction site.
up+ or upoconstants (13). These studies on the chemical mechanism of the enzyme
may also be useful in explaining the dead end inhibition
DISCUSSION caused by certain substituted phenols. Those phenols with
Aryl sulfotransferase IV from rat liver catalyzes three types several strongly electron-withdrawing substituents, e.g. pen-
of reactions: the sulfation of phenols in which PAPS serves as tachlorophenol and 2,6-dichloro-4-nitrophenol(17), are excel-
donor (Equation 1);the PAP-mediated sulfation of a phenol lent dead end inhibitors. Although there is a steric contribu-
by a phenyl sulfate, the “exchange reaction” (Equation 2); tion due to substituents in the 2 and 6 position^,^ this is not
andthe nucleotide-independent hydrolytic cleavage of a sufficient for complete inhibition of sulfation. Rather, elec-
phenyl sulfate (Equation 5). tron-withdrawing substituents decrease the rate of sulfation
Based on akinetic analysis that includes patterns of product There is a 40% difference in the rate of sulfation between 2,4-
inhibition, the kinetic mechanism of the overall reaction dimethylphenol and 2,6-dimethylphenol when measured at a concen-
(Equation 3) is consistent with either arapid equilibrium tration of 100 p ~ pH , 7.0, 25 “C, with 200 PM PAPS.
 Mechanism
SulfotransferaseAryl
without decreasing the affinity of the substituted phenol for
the enzyme. Pentachlorophenol, for example, inhibits sulfa-
tion of CNP with a K , of 0.2 p ~ These
. effects of electron-
withdrawing substituents may prove useful in designingboth
inhibitors and substrates for this aryl sulfotransferase.
Acknowledgment-We are grateful to John Westley for a critical
discussion of the work.
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