C-terminal deletion analogs of a crustacean pigment-dispersing hormone

This study deals with the effect of deamidation and C-terminal truncation on the potency of an octadecapeptide pigment-dispersing hormone (PDH: Asn- Ser-Gly-Met-Ile-Asn-Ser-Ile-Leu-Gly-Ile-Pro-Arg-Val-Met-Thr-Glu-Ala-NH₂), first described as light-adapting distal retinal pigment hormone (DRPH) from Pandalus borealis. Bioassay of synthetic analogs for melanophore pigment dispersion in destalked fiddler crabs (Uca pugilator) showed that deamidation causes a 300-fold decrease in potency. The analogs 1–17-NH₂ and 1–16-NH₂ were about 3 times more potent than 1–18-OH. Further truncation led to decreases in potency, with the peptide 1–9-NH₂ being the smallest C-terminal deletion analog to display activity (0.001% potency). Smaller analogs (1–8-NH₂, 1–6-NH₂ and 1–4-NH₂) were inactive when tested in doses as high as 500 nmoles/crab. On the basis of our earlier work on N-terminal deletion analogs and the present findings the residues 6 to 9 seem to be important for PDH action.     

Characterization of growth hormone-releasing hormone (GHRH) binding to cloned porcine GHRH receptor

To study structure-activity relationships of growth hormone-releasing hormone (GHRH), a competitive binding assay was developed using cloned porcine adenopituitary GHRH receptors expressed in human kidney 293 cells. Specific binding of [His¹,¹²⁵I-Tyr¹⁰,Nle²⁷]hGHRH(1–32)-NH₂ increased linearly with protein concentration (10–45 μg protein/tube). Binding reached equilibrium after 90 min at 30°C and remained constant for at least 240 min. Binding was reversible to one class of high-affinity sites (K_d = 1.04 ± 0.19 nM, B_max = 3.9 ± 0.53 pmol/mg protein). Binding was selective with a rank order of affinity (IC₅₀) for porcine GHRH (2.8 ± 0.51 nM), rat GHRH (3.1 ± 0.69 nM), [N-Ac-Tyr¹, -Arg²]hGHRH(3–29)-NH₂ (3.9 ± 0.58 nM), and [ -Thr⁷]GHRH(1–29)-NH₂ (189.7 ± 14.3 nM), consistent with their binding to a GHRH receptor. Nonhydrolyzable guanine nucleotides inhibited binding. These data describe a selective and reliable method for a competitive GHRH binding assay that for the first time utilizes rapid filtration to terminate the binding assay.     

Novel method of enzymes stabilization on crosslinked thermosensitive carriers

In this paper an immobilization of invertase on thermosensitive copolymers of N-isopropylacrylamide and 2-hydroxyethyl methacrylate or glycidyl methacrylate modified by aminolysis is evaluated. The method is based on the swelling properties of stimuli-sensitive polymers, which work like a pump that sucks up the enzyme on cooling and then on subsequent crosslinking of the enzyme. The attention was focused on the properties of the carrier–enzyme systems, particularly on the effect of crosslinking on their stability. Activity of TG8-NH₂ carrier was very low and independent on concentration of glutaraldehyde used, but carriers TH8 and TH8-NH₂ were more active, especially when 1.0 and 2.5 vol.% of glutaraldehyde were used. It was also observed, that preparations crosslinked by glutaraldehyde were more stable than preparations without crosslinking agent.     

High affinity receptors for vasoactive intestinal peptide on a human glioma cell line

Vasoactive intestinal peptide (VIP) bound with high affinity (K_d 0.13 nmol/l) to receptors on the human glioma cell line U-343 MG Cl 2:6. The receptors bound the related peptides helodermin. PHM and secretin with 10, 400 and 5000 times lower affinity, respectively. Deamidated VIP (VIP-COOH) and [des-His¹]VIP bound with 10 and 100 times lower affinity. The fragment VIP(7–28) displaced 25% of the receptor-bound ¹²⁵I-VIP whereas VIP(16–28) and VIP(1–22-NH₂) were inactive. The binding of ¹²⁵I-VIP could be completely inhibited by 10 μmol/l of the antagonists [N-Ac-Tyr¹,D-Phe²]GRF(1–29)-NH₂, [pCl-D-Phe⁶,Leu¹⁷]VIP and VIP(10–28); in contrast, the antagonist L-8-K was inactive. Affinity labeling showed that VIP bound to proteins with Mr's of 75 kDa, 66 kDa and 50 kDa, respectively. Following binding, the peptide was rapidly internalized, and at steady-state only 20% of cell-associated ¹²⁵I-VIP was bound to receptors on the cell surface. The internalized ¹²⁵I-VIP was completely degraded to ¹²⁵I-tyrosine which was released from the cells. Degradation of internalized ¹²⁵I-VIP was significantly reduced by chloroquine phenantroline and pepstatin-A. Surface binding and internalization of ¹²⁵I-VIP was increased 3 times by phenantroline, and pepstatin-A caused a 5 times increase in surface binding. Chloroquine reduced surface-bound ¹²⁵I-VIP, but caused retention of internalized ¹²⁵I-VIP.     

Lack of effect of vasoactive intestinal peptide antagonists on blood flow in the rat thyroid

We used three putative vasoactive intestinal peptide (VIP) antagonists: 1) [4Cl-D-Phe⁶,Leu¹⁷]VIP, 2) [N-Ac-Tyr¹,D-Phe²]GRF(1–29)-NH₂, and 3) VIP(10–28) to assess the involvement of endogenous VIP in the regulation of thyroid hormone secretion and thyroid blood flow (BF). We measured thyroid BF in ketamine-pentobarbital-anesthetized rats using the microsphere technique. Increases in thyroid BF induced by VIP administration (30 pmol-1.5 nmol/100 g b.wt.) were not affected by any of the three compounds tested at doses 10–100 times higher than that of VIP. These compounds (3–15 nmol/100 g b.wt.) also failed to affect basal thyroid BF or hormone secretion. Increases in pancreatic and salivary gland BFs induced by VIP (30 pmol/100 g b.wt.) were also not affected by [4Cl-D-Phe⁶,Leu¹⁷]VIP or [N-Ac-Tyr¹,D-Phe²]GRF(1–29)-NH₂ (3 nmol/100 g b.wt.). These results indicate that the three compounds tested are not effective inhibitors of VIP receptors in the thyroid vasculature and, therefore, they cannot be used in the investigation of the functional significance of endogenous VIP in the regulation of thyroid BF.     

Exoprotease activity of Leuconostoc oenos in stress conditions

Exoprotease activity during 48 h of total energy and nutrient starvation was examined in Leuconostoc oenos X₂L isolated from wine. Starved cells after 2 h of incubation at 30 °C in citrate buffer, 0.05 mmol 1⁻¹ pH 5, showed greater extracellular proteolytic activity than at the onset of starvation. In the presence of 60 mg l⁻¹ SO₂ and 8% or 12% ethanol, the proteolytic activity was higher ; 10 mmol l⁻¹ Ca²⁺ and Mg²⁺ produced an increase in protease activity during starvation. Glucose and 2-deoxyglucose (2-DOG) were found to repress synthesis by 80% and 100%, respectively. Cyclic adenosine 3'-5'-phosphate increased the exoprotease activity and reverted the repression by glucose and 2-DOG. De novo synthesis of proteins was required for the exoprotease activity by cells submitted to stress conditions. The absence of protease activity in the supernatant fluids from chloramphenicol-treated cells indicated that the activity is a result of deliberate release and not of passive cell lysis.     

Chimeric NDP-MSH and MTII melanocortin peptides with agouti-related protein (AGRP) Arg-Phe-Phe amino acids possess agonist melanocortin receptor activity

Agouti-related protein (AGRP) is one of only two known endogenous antagonists of G-protein coupled receptors (GPCRs). Specifically, AGRP antagonizes the brain melanocortin-3 and -4 receptors involved in energy homeostasis, regulation of feeding behavior, and obesity. -Melanocyte stimulating hormone (-MSH) is one of the known endogenous agonists for these receptors. It has been hypothesized that the Arg-Phe-Phe (111–113) human AGRP amino acids may be mimicking the melanocortin agonist Phe-Arg-Trp (7–9) residue interactions with the melanocortin receptors that are important for both receptor molecular recognition and stimulation. To test this hypothesis, we generated thirteen chimeric peptide ligands based upon the melanocortin agonist peptides NDP-MSH (Ac-Ser-Tyr-Ser-Nle⁴-Glu-His-DPhe-Arg-Trp-Gly-Lys-Pro-Val-NH₂) and MTII (Ac-Nle-c[Asp-His-DPhe-Arg-Trp-Lys]-NH₂). In these chimeric ligands, the agonist DPhe-Arg-Trp amino acids were replaced by the AGRP Arg-Phe-Phe residues, and resulted in agonist activity at the mouse melanocortin receptors (mMC1R and mMC3–5Rs), supporting the hypothesis that the AGRP antagonist ligand Arg-Phe-Phe residues mimic the agonist Phe-Arg-Trp amino acids. Interestingly, the Ac-Ser-Tyr-Ser-Nle⁴-Glu-His-Arg-DPhe-Phe-Gly-Lys-Pro-Val-NH₂ peptide possessed 7 nM mMC1R agonist potency, and is 850-fold selective for the mMC1R versus the mMC3R, 2300-fold selective for the mMC1R versus the mMC4R, and 60-fold selective for the MC1R versus the mMC5R, resulting in the discovery of a new peptide template for the design of melanocortin receptor selective ligands.     

Phospholipase A2 in human placental serum

Intervillous blood was collected from term placentae at delivery, and sera were tested for phospholipase A₂ under various experimental conditions. Enzyme activity was found to develop upon extended storage in the cold or at 37°C. The enzyme is reversibly inhibited by dithiothreitol, requires Ca⁺⁺ ions for activity, and tolerates various detergents. The apparent molecular weight is 42 kDa. In all these parameters the serum enzyme behaves similar to the 42 kDa phospholipase A₂ which we recently purified to homogeneity from thoroughly washed placental tissue. Serum phospholipase A₂ appears to be generated by proteolytic processing from a slightly larger inactive precursor which was detected immunochemically. Most likely this protein originates from fetal cells and may be released by membrane damage. We conclude that both placental serum and tissue harbour a novel type of phospholipase A₂ which is distinct from cytosolic and secretory phospholipases A₂. Preference for arachidonate containing substrate suggests a role in eicosanoid production within gestational tissues.     

Immunosensor for the detection of Vibrio cholerae O1 using surface plasmon resonance

An immunosensor for the detection of Vibrio cholerae O1 was developed on the basis of surface plasmon resonance (SPR). A protein G layer was fabricated by means of the chemical coupling between the free amine (-NH₂) groups of protein G and the activated carboxyl groups present on a self-assembled monolayer (SAM) consisting of a mixture of 11-mercaptoundecanoic acid (MUA) and hexanethiol (molar ratio of 1:2). A monoclonal antibody, which was confirmed to be specific to V. cholera O1 by the Western blotting technique, was immobilized on the protein G layer. The formation of the SAM, the protein G layer and the sequential binding of the antibody against V. cholera O1 were investigated with SPR spectroscopy. As the number of fabricated layers increased, the minimum angle of plasmon resonance was increased accordingly. The target bacteria, V. cholera O1, was measured with the fabricated immunosensor, whose detection range was between 10⁵ and 10⁹ cells/mL.     

Studies of the extracellular proteolytic activity produced by Propionibacterium acnes

Of several commercial media tested, trypticase soya broth containing 0.4% (w/v) D-sorbitol was superior as a growth medium for the production of extracellular proteinase by Propionibacterium acnes (strain P-37). Extracellular proteinase, production of which was shown to be growth-associated by both batch and continuous culture studies, was partially purified by 70% (NH₄)₂SO₄ saturation, Sephadex G-75 chromatography and ion exchange on DEAE-Sephadex A-50. It was shown to be a heterogeneous mixture of at least three molecular species of enzyme. Proteinase I was inhibited by EDTA (10^-3 mol/l) and PMSF (5 millimol/l) and stimulated by CaCl₂ (190% at 10^-3 mol/l). It had a molecular weight of 20 to 30000 and a broad pH optimum from 6.5 to 7.5. Proteinase II was an alkaline proteinase with a molecular weight of 30 to 40000 which was not significantly inhibited by EDTA (10^-2 mol/l) nor stimulated by CaCl₂. Proteinase III represented a minor proportion of the recovered proteolytic activity, had a molecular weight of 20 to 30000 and was most active in the alkaline pH range. This enzyme was inhibited by EDTA (10^-4 mol/l) and PMSF (5 millimol/l), and stimulated by CaCl₂ (250% at 10^-2 mol/l).     

The influence of anionic lipids on SHIP2 phosphatidylinositol 3,4,5-trisphosphate 5-phosphatase activity

The SH2 domain containing inositol 5-phosphatase 2 (SHIP2) catalyzes the dephosphorylation of phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P₃) to phosphatidylinositol 3,4-bisphosphate (PtdIns(3,4)P₂) and participates in the insulin signalling pathway in vivo. In a comparative study of SHIP2 and the phosphatase and tensin homologue deleted on chromosome 10 (PTEN), we found that their lipid phosphatase activity was influenced by the presence of vesicles of phosphatidylserine (PtdSer). SHIP2 PtdIns(3,4,5)P₃ 5-phosphatase activity was greatly stimulated in the presence of vesicles of PtdSer. This effect appears to be specific for di-C8 and di-C16 fatty acids of PtdIns(3,4,5)P₃ as substrate. It was not observed with inositol 1,3,4,5-tetrakisphosphate (Ins(1,3,4,5)P₄) another in vitro substrate of SHIP2, nor with Type I Ins(1,4,5)P₃/Ins(1,3,4,5)P₄ 5-phosphatase activity, an enzyme which acts on soluble inositol phosphates. Vesicles of phosphatidylcholine (PtdCho) stimulated only twofold PtdIns(3,4,5)P₃ 5-phosphatase activity of SHIP2. Both a minimal catalytic construct and the full length SHIP2 were sensitive to the stimulation by PtdSer. In contrast, PtdIns(3,4,5)P₃ 5-phosphatase activity of the Skeletal muscle and Kidney enriched Inositol Phosphatase (SKIP), another member of the mammaliam Type II phosphoinositide 5-phosphatases, was not sensitive to PtdSer. Our enzymatic data establish a specificity in the control of SHIP2 lipid phosphatase activity with PtdIns(3,4,5)P₃ as substrate which is depending on the fatty acid composition of the substrate.     

大熊猫尸体组织LDH同工酶盘电泳观察

大熊猫是冰川时期残存至今的古老、稀有的珍贵动物,有“活化石”之称。对大熊猫的研究除野外调查(王朗自然保护区大熊猫调查组,1974)、形态解剖(张鹤宇等,1959;冯文和等,1984)、人工饲养繁殖(北京动物园,1974;冯文和等,1983;1984)等外,尚有生化技术和免疫学等研究方法(Sarich,1973;潘文石等,1982;罗昌容等,1984)。     

An N-Protected γ-Aminobutyric Acid Dipeptide with Anticonvulsant Action

Abstract: N -Pivaloyl-leucyl–γ-aminobutyric acid (PLG) is a synthetic dipeptide with a partition coefficient of 1.67 in an ethyl acetate/water system that partially inhibits the synaptosomal uptake and activates the release of [U- ¹⁴C]-γ-aminobutyric acid ([U-¹⁴C]GABA). The displacement of GAB A from crude synaptic membranes by PLG occurs with an IC₅₀ of 10⁻⁵ M . The compound has the capacity to cross the blood-brain barrier and increase central GABA levels. Its ED₅₀ on cardiazol-induced convulsions is 60-65 mg/kg. PLG is resistant to hydrolysis by chymotrypsin and partially inhibits the proteolytic activity of trypsin.     

胞外三磷酸腺苷对铅胁迫下植物细胞损伤和过氧化氢及其清除酶水平的调节

细胞外三磷酸腺苷（extracellular adenosine-5'-triphosphate）是植物细胞的重要信号分子。以烟草悬浮细胞BY-2（Nicotiana tabacum L.cv.Bright Yellow-2）为材料,探讨了胞外三磷酸腺苷对铅胁迫下细胞损伤、H₂O₂（过氧化氢）含量及H₂O₂清除酶活性的影响。结果显示,随着Pb（NO₃）₂浓度的不断提高（30~400 μmol·L^-1）,细胞外三磷酸腺苷含量呈现出逐渐下降的趋势,但胞内三磷酸腺苷含量及细胞的受损伤程度逐渐增大;同时,H₂O₂含量和过氧化氢酶的活性均有所上升,并在200 μmol·L^-1 Pb（NO₃）₂处理下达到最大值,而过氧化物酶的活性则不断降低。较之Pb（NO₃）₂胁迫下的细胞,对Pb（NO₃）₂胁迫的细胞加入外源三磷酸腺苷使得细胞受损伤程度显著降低,H₂O₂含量减少,过氧化氢酶活性减弱,而过氧化物酶活性增强。实验结果表明,Pb（NO₃）₂胁迫诱导的植物细胞损伤和H₂O₂及其清除酶水平的变化能受到细胞外三磷酸腺苷水平的调节。     

Effects of different gibberellins on elongation growth under short day conditions in seedlings of Salix pentandra

Fifteen different gibberellins (GA's) were tested for their ability to induce elongation growth under short day conditions in seedlings of Salix pentandra L. GA's were applied either to the apex or they were injected into a mature leaf. GA₃ was highly active and also GA₄₊₇ and GA₄ showed high activity. GA₁, GA₂, GA₅, GA₉, GA₁₃, GA₂₀, GA₃₆ and GA₄₇ showed moderate activity. GA₁₆, GA₁₇, GA₂₇ and GA₄₁ exhibited low or no activity in doses up to 10 μg per plant. In general, a better growth response was obtained with an application to the apex than with an injection into the leaf.     

Characterization of phospholipase A2 and acyltransferase activities in squid (Loligo pealei) axoplasm: comparison with enzyme activities in other neural tissues, axolemma and axoplasmic subfractions.

Phospholipase A₂ and acyltransferase were assayed and characterized in pure axoplasm and neural tissues of squid. Intracellular phospholipase A₂ activity was highest in giant fiber lobe and axoplasm, followed by homogenates from retinal fibers, optic lobe and fin nerve. In most preparations, exogenous calcium (5 mM) caused a slight stimulation of activity. EGTA (2 mM) was somewhat inhibitory, indicating that low levels of endogenous calcium may be required for optimum activity. Phospholipase A₂ was inhibited by 0.1 mM p-bromophenacylbromide, and was completely inactivated following heating.

The level of acylCoA: lysophosphatidylcholine acyltransferase activity was higher in axoplasm and giant fiber lobe than in other neural tissues of the squid. K_m (apparent) and V_max (apparent) for oleoyl-CoA and lysophosphatidylcholine were quite similar for axoplasm and giant fiber lobe enzyme preparations. Acyltransferase activity was inactivated by heat treatment, and greatly inhibited by 0.2 mM p-chloromercuribenzoate, and to a lesser extent by 20 mM N-ethylmaleimide.

Phospholipase A₂ activity was present in fractions enriched in axolemmal membranes (separated from squid retinal fibers and garfish olfactory nerve) from both tissues, and it was also highly concentrated in vesicles derived from squid axoplasm. In all three preparations, phospholipase A₂ activity was stimulated by Ca⁺⁺ (5 mM) and inhibited by EGTA (2 mM). In addition, axoplasmic cytosol (114,000 g supernatant) retained a substantial portion of a Ca⁺⁺-independent phospholipase A₂, active in the presence of 2 mM EGTA. Acyltransferase activity was present at high content in both axolemma membrane rich fractions, and among subaxoplasmic fractions and axoplasmic vesicles.     

Effect of carbon dioxide on antioxidant enzymes and ginsenoside production in root suspension cultures of Panax ginseng

The aim of this study was to investigate the effect of CO₂ at various concentrations (1, 2.5 and 5%) on antioxidant enzymes and ginsenoside accumulation in Panax ginseng roots in 5 l airlift bioreactors (working volume 4 l). One and 2.5% CO₂ was beneficial for root biomass accumulation, but 5% CO₂ decreased the biomass. Ginsenoside concentration decreased with increasing concentration of CO₂. No significant difference was observed in the malondialdehyde (MDA) content and lipoxygenase (LOX) activity between respective controls and CO₂ treated roots. Antioxidant enzymes such as ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), glutathione reductase (GR), catalase (CAT), guaiacol peroxidase (G-POD) including reduced ascorbate and total glutathione were induced in CO₂ exposed roots which emphasized the protective role of antioxidants against CO₂ induced stress. Superoxide dismutase activity (SOD) which was induced after 15 days was significantly inhibited after 45 days. Glutathione-S-transferase (GST) and glutathione peroxidase (GPX) activities also increased when the roots were subjected to 1 and 2.5% CO₂ compared to the respective controls but not at 5%. A higher reduced ascorbate to oxidized (ASC/DHA) ratio in CO₂ treated root indicates the plant's ability to tolerate CO₂ stress. These observations suggest that an increase in antioxidant enzymes may affect a defense response to the cellular damage induced by CO₂. Probably, this increase could not stop the deleterious effects of CO₂ concentration on ginsenoside concentration, but reduced stress severity and thereby allowing root growth to occur.     

解脂假丝酵母对重金属工业废水的吸附特性

利用解脂假丝酵母对Cr(Ⅵ)、Ni(Ⅱ)和Cu(Ⅱ)共存的模拟重金属废水及3种实际重金属废水进行了微生物吸附,结果表明,pH、吸附时间和菌浓度等均是显著的影响因素.Cr(Ⅵ)、Ni(Ⅱ)和Cu(Ⅱ)的去除均符合准一级和准二级动力学模型,其中准二级模型的拟合效果最理想,证明该菌种对重金属的吸附包括了多个步骤,其中化学吸附是限速步骤.解脂假丝酵母对共存重金属的生物吸附效果理想,1g·L^-1菌体在120min时,对18.7～37.86mg·L^-1Cr、2.39～9.21mg·L^-1Cu、2.27～9.87mg·L^-1Ni和0.43～1.32mg·L^-1Zn的去除率分别为81.6%～84.6%、84.0%～100%、84.1%～100%和93.9%～100%.菌体的蛋白质、脂质和多糖均参与了重金属吸附,起作用的主要功能团是-OH、-NH₂、-CH₂、-CH₃、-COOH、-CHO、C=C、-PO₄^3-和-SO₃H.     

Water stress, carbon dioxide, and light effects on sucrosephosphate synthase activity in Phaseolus vulgaris

The characteristics of sucrose-phosphate synthase (SPS; EC 2.4.1.14) activity in leaves of Phaseolus vulgaris L. cv. Linden was studied in plants subjected to water stress and various CO₂ and light treatments. When water was withheld for 3 days causing mild water stress (–0.9 MPa), the activity of SPS measured in crude extracts was reduced ca 50%. The effect of water stress was most evident when the enzyme was assayed with saturating amounts of its substrates fructose 6-phosphate and UDP glucose. Placing a water-stressed plant in an atmosphere containing 1% CO₂ reversed the effect of water stress on SPS activity over 5 h even though the water stress was not relieved. Holding unstressed leaves in low CO₂ partial pressure reduced the extractable activity of SPS. After 1 h of low CO₂ treatment the effect of low CO₂ could be reversed by 20 min of 5% CO₂. However, after 24 h of low CO₂ treatment, less SPS activity was recovered by the 20 min treatment. The cytosolic protein synthesis inhibitor cycloheximide prevented the slow recovery of SPS activity, but did not affect the rapid recovery of SPS. We conclude that the effect of water stress on SPS activity was a consequence of the inhibition of photosynthesis caused by stomatal closure. Responses of Phaseolus vulgaris SPS to light were similar to the response to low CO₂ in that the effects were most pronounced under V_max assay conditions. This is the first report of this type of light response of SPS in a dicotyledonous species.     

Effect of SO2, on the activity of adenosine 5'-phosphosulfate sulfotransferase from spruce trees (Picea abies) in fumigation chambers and under field conditions

The effect of SO₂ on adenosine 5'-phosphosulfate sulfotransferase activity and various other parameters of needles from spruce ( Picea abies L.) was studied using potted grafts in outdoor fumigation chambers and trees growing near a factory. In summer and autumn fumigation of grafted spruce, SO₂, decreased the extractable activity of adenosine 5'-phosphosulfate sulfotransferase to 12–50% of the controls, and reduced the amount of ³⁵S from sulphate incorporated into protein by excised branches to a comparable degree. SO₂ treatment in January and February inhibited the increase in adenosine 5'phosphosulfate sulfotransferase activity measured in the controls during this time. ATP-sulfurylase activity was less affected by SO₂. fumigation. In trees growing near a factory with high SO₂. emission, the activity of adenosine 5'-phosphosulfate sulfotransferase was about 35% of that of trees from a control area. The low enzyme activity was correlated with a high content of sulfate and compounds containing thiol groups.