Human Platelet Phenol Sulfotransferase: Partial Purification and Detection of Two Forms of the Enzyme

To begin to study the usefulness of platelet phenol sulfotransferase (PST) as a possible measure of the enzyme activity in other organs such as the brain, we purified human platelet PST 36-120-fold. Activity toward 3-methoxy-4-hydroxyphenylglycol (MHPG), dopamine, 5-hydroxytryptamine (5-HT), and phenol eluted in the same Sephadex G-100 and Affi-Gel Blue column fractions. Specific activities of the enzyme with MHPG, dopamine, 5-HT, and phenol as substrates were 1198, 1068, 401, and 408 units/mg protein, respectively. Optimal assay conditions were established for each substrate. Apparent Km values were 598 microM, 21 microM, 19 microM, and 500 microM for MHPG, dopamine, phenol, and 5-HT, respectively. Apparent Km values for 3'-phosphoadenosine-5'-phosphosulfate (PAPS) with the same four substrates ranged from 0.11 to 0.25 microM. The pH optima were 6.3 for phenol, 6.8 for dopamine, and 7.0 for MHPG and 5-HT. An additional pH optimum at 8.6 was present for 5-HT. A thermolabile form of the enzyme measured with dopamine and 5-HT, as well as a thermostable form measured with phenol, were present. Dichloronitrophenol (10(-5) M) noncompetitively inhibited the thermostable enzyme activity by 96% but decreased the thermolabile activity by only 36%. These studies provide the basis for a more accurate comparison of human platelet PST with the enzyme in the human brain and in other tissues.     

Human Phenol Sulfotransferase: Correlation of Brain and Platelet Activities

Phenol sulfotransferase (PST; EC 2.8.2.1) catalyzes the sulfate conjugation of phenolic and catechol neurotransmitters and drugs. The human blood platelet has been the most thoroughly studied source of PST because of the possibility that the regulation of the enzyme in this easily accessible tissue might reflect the regulation of PST in the CNS. The human brain and platelet contain at least two forms of PST, forms designated as thermostable (TS) and thermolabile (TL) PST. TS PST catalyzes the sulfate conjugation of micromolar concentrations of phenol and p-nitrophenol and TL PST catalyzes the sulfate conjugation of dopamine and other monoamines. This study was performed to determine whether individual variations in the activities of human platelet TS and TL PST reflect individual variations in cerebral cortical PST activities. PST activities were measured in platelets and in cerebral cortical tissue obtained from 15 patients with epilepsy during clinically indicated neurosurgery. There was a highly significant correlation between the activities of the TS form of PST in cerebral cortex and platelets of these patients (r = 0.940, p less than 0.001), but there was not a significant correlation between activities of the TL form of PST in the two tissues (r = 0.396, p greater than 0.14). In addition to variations in the level of enzyme activity, there are also wide individual variations in the thermal stability of platelet TS PST.(ABSTRACT TRUNCATED AT 250 WORDS)     

Human Brain Phenol Sulfotransferase: Biochemical Properties and Regional Localization

Phenol sulfotransferase (PST) catalyzes the sulfate conjugation of catecholamines and phenol and catechol drugs. The human blood platelet contains a thermolabile (TL) form of PST that catalyzes the sulfate conjugation of dopamine and other monoamines and a thermostable (TS) form that catalyzes the sulfate conjugation of micromolar concentrations of phenol and p-nitrophenol. Experiments were performed to determine whether the brain contains forms of PST analogous to the TL and TS forms found in the human platelet, and to determine whether there are regional variations in human brain PST activity. We found that the human brain contains at least two forms of PST, forms that are similar to the platelet TS and TL forms of the enzyme with respect to substrate specificity, apparent Km constants, thermal stability, and sensitivity to inhibitors. Optimal conditions were determined for the measurement of these two activities in brain homogenates. The stability of PST activities in the human brain after death was determined in five samples of cerebral cortex that were obtained during clinically indicated neurosurgical procedures. An average of 76 +/- 8% and 80 +/- 9% (mean +/- SEM) of the basal TL and TS PST activities, respectively, remained in these five samples of cerebral cortex after 8 h of storage under simulated post-mortem conditions. Six human brains were then obtained less that 8 h after death from patients who had no neurological disease prior to death. The mean activities of the TL and TS forms of PST were measured in 17 different regions of the six brains. If the pituitary was excluded from consideration, TL and TS PST activities both varied approximately fivefold among these regions, and both activities were highest in cerebral cortex. However, the average TS activity in the anterior pituitary, a tissue of non-neural origin embryologically, was 6.5-fold greater than the highest average TS PST activity found in cerebral cortex.     

大鼠肾硫酸基转移酶基因的cDNA克隆及序列分析

在许多激素、神经递质、药物和异生化合物的生物转化中,硫酸酯化反应是一重要的代谢途径［１,２］．这些化合物的硫酸酯化通常导致其生物活性的降低及尿排泄量的增加．硫酸酯化是把３′－磷酸腺苷－５′磷酸硫酸（ＰＡＰＳ）的活性硫酸根转移到某一底物（如Ｒ－ＯＨ）．...     

重庆赤车属一新种和楼梯草属二新种

描述了自重庆市江津区四面山发现的荨麻科赤车属一新种和楼梯草属二新种：（1）四面山赤车,与融安赤车近缘,区别在于新种的叶的二级脉较少,2枚雌花被片有短角状突起,以及长圆形瘦果;（2）稀柱毛楼梯草,可能与托叶楼梯草近缘,区别在于新种的叶较小,雌总苞苞片,以及雄、雌小苞片均在背面具龙骨状突起,柱头由只3~4根毛组成;（3）四面山楼梯草,与长尖楼梯草近缘,区别在于新种茎的顶端被短柔毛,雄头状花序的花序梗较短,以及雄总苞苞片呈条状船形,背面具龙骨状突起。     

Sulfation of Dopamine and Other Biogenic Amines by Human Brain Phenol Sulfotransferase

Abstract: Phenol sulfotransferase was isolated in 100,000g supernatant fractions prepared from postmortem samples of human brain. Since phenol sulfotransferase (PST) has been shown to conjugate the amine neurotransmit-ters in vivo , the abilities of eight different biogenic amines and structurally related compounds to act as substrates for PST were studied. These experiments demonstrate that at a concentration of 20 μM, dopamine (DA) was the best substrate examined and was followed in decreasing order of activity by 3-methoxytyramine (3-MT), tyramine, norepinephrine, 3-methoxy-4-hydroxyphenylethyleneglycol, octopamine, 5-hydroxytryptamine and dihydroxyphenylethyleneglycol. At a substrate concentration of 100 /UM the relative order of activity was altered, so that tyramine became the most rapidly conjugated substrate while the activity of DA and 3-MT relative to the other substrates tested was diminished. This change in substrate affinity with differing substrate concentrations can be explained, at least for DA, by the occurrence of apparent substrate inhibition at concentrations above 25 to 30 μM. Using PST isolated in 100,000g supernatant fractions from human brain, the K_m value for DA was found to be 5.0 μM, while the K_m value for the sulfate-donor 3'-phosphoadenosine-5'-phosphosulfate was 0.25 μM. The ratio of 3- O - to 4- O -DA-sulfate formed in vitro by human brain PST was found to be about 4: 1. In addition, both the 3- O - and 4- O -esters were found not to be deaminated by human brain mitochondrial MAO. The relative role of PST with respect to MAO and catechol- O -methyltransferase in the degradation of the biogenic amine neurotransmitters in human brain is discussed.     

中国尖翅蝇属二新种及一新记录种记述(双翅目:尖翅蝇科)

记述采自中国北方尖翅蝇属Lonchoptera Meigen 2新种,即纤毛尖翅蝇L.ciliosa,sp.nov.和多鬃尖翅蝇L.multiseta,sp.nov.,讨论了它们与近缘种的区别.并首次记录了亮额尖翅蝇L.nitidifrons Strbol在中国的分布.模式标本保存在中国农业大学昆虫博物馆.     

中国园蛛科1新种和2个新雄性(蜘蛛目:园蛛科)

本文描述了园蛛科3种蜘蛛：多齿园蛛,新种Araneus polydentatus sp．nov．,舜皇园蛛Araneus shunhuangensis Yin et al,1990和松阳芒果蛛Mangora songyangensis Yin et al,1990。舜皇园蛛和松阳芒果蛛的雄性均系首次报道。本文同时提供了这3种蜘蛛的地理分布图。     

Corticosteroid-Binding Globulin: Structure-Function Implications from Species Differences

Corticosteroid-binding globulin (CBG) transports glucocorticoids and progesterone in the blood and thereby modulates the tissue availability of these hormones. As a member of the serine protease inhibitor (SERPIN) family, CBG displays a reactive center loop (RCL) that is targeted by proteinases. Cleavage of the RCL is thought to trigger a SERPIN-typical stressed-to-relaxed (S-to-R) transition that leads to marked structural rearrangements and a reduced steroid-binding affinity. To characterize structure-function relationships in CBG we studied various conformational states of E. coli-produced rat and human CBG. In the 2.5 Å crystal structure of human CBG in complex with progesterone, the RCL is cleaved at a novel site that differs from the known human neutrophil elastase recognition site. Although the cleaved RCL segment is five residues longer than anticipated, it becomes an integral part of β-sheet A as a result of the S-to-R transition. The atomic interactions observed between progesterone and CBG explain the lower affinity of progesterone in comparison to corticosteroids. Surprisingly, CD measurements in combination with thermal unfolding experiments show that rat CBG fails to undergo an S-to-R transition upon proteolytic cleavage of the RCL hinting that the S-to-R transition observed in human CBG is not a prerequisite for CBG function in rat. This observation cautions against drawing general conclusions about molecular mechanisms by comparing and merging structural data from different species.     

中国夜蛾科一新纪录属和二新纪录种记述(鳞翅目)

通过整理近年采自西藏自治区色季拉山的夜蛾科Noctuidae标本材料,发现夜蛾科一中国新纪录属:耳夜蛾属Dioszeghyana Hreblay,1993和二中国新纪录种:黑双耳夜蛾Dioszeghyana nigralba(Yoshimoto,1993)和同灰夜蛾Polia similissima Plante,1982。对所研究的物种给予了详细的外部形态特征和外生殖器描述,并提供了其国内外分布信息。     

侧耳属(Pleurotus)二个野生种与一个栽培种之间亲和性研究

用从校内采集的P 96 6和从德阳市采集的P DY97两个侧耳属野生种与栽培种P 17之间的单核体进行配对具有不亲和性 ,证明了这 3个物种间遗传的独立性。同时通过P 17,P 96 6 ,P DY97各自的单核体自交实验 ,表明这 3个物种的交配系统是双因子控制系统。     

Inhibition of M and P Phenol Sulfotransferase by Analogues of 3''-Phosphoadenosine-5''-Phosphosulfate

Structural analogues of the sulfate donor 3'-phosphoadenosine-5'-phosphosulfate (3',5'-PAPS) were examined for their ability to inhibit dopamine and phenol sulfation by the M and P forms of phenol sulfotransferase (PST), respectively. The Ki values for each of the adenosine derivatives were calculated from the rate equation for PST. For both M and P PST, the naturally occurring product 3'-phosphoadenosine-5'-phosphate, (3',5'-PAP), was shown to be the most effective inhibitor. The weakest inhibitors of the two sulfotransferases were 5'-adenosine phosphosulfate and the three AMP derivatives, which were less than 1,000 times as effective as 3',5'-PAP. 5'-ATP, 2',5'-PAPS, 2',5'-PAP, and 5'-ADP were similar in their inhibition of M and P PST and were all approximately 100 times less effective than the natural end product. These data reveal that there is a rigid structural requirement for binding of the ribose portion of adenosine to both M and P PST that involves the groups on both the 3' and 5' positions. The effectiveness of binding to the two enzymes may depend on both steric factors as well as the distribution of negative charges on the ribose ring.     

广东省车八岭自然保护区连唇步甲属一新种记述（鞘翅目：步甲科）

记述采自中国广东始兴车八岭国家级自然保护区连唇步甲属Ｓｏｆｏｔａ１新种：黑连唇步甲Ｓｏｆｏｔａ ｎｉｇｒｕｍ,ｓｐ．ｍｏｖ．。模式标本保存在华南农业大学昆虫标本室。     

相异物种同源基因cDNA的快速克隆

介绍了一种结合生物信息学数据库的比较分析,在系统发生相近物种的棱酸保守区设计PCR引物,通过RT-PCR和RACE方法,在相异物种中快速克隆同源基因cDNA的一种简捷方法。     

中国狼逍遥蛛属1新种2新纪录种记述（蜘蛛目：逍遥蛛科）

记述分布于内蒙古的逍遥蛛科蜘蛛1新种:武川狼逍遥蛛Thannatus wuchuanensis sp.nov和2中国新纪录种:北极狼逍遥蛛Thanatus arctivus Thorell,1872;草原狼逍遥蛛Thanatus stepposus Logunov,1996.     

两种丝状绿藻对水体中低浓度苯酚的去除作用研究

研究了两种丝状绿藻,脆弱刚毛藻(Cladophora fracta)和长形水绵(Spirogyra longata)对水体中低浓度苯酚(≤6mg·mL-1)的去除作用。结果表明,在不考虑交互作用下,脆弱刚毛藻和长形水绵对苯酚的去除率分别为86%和75.63%,脆弱刚毛藻去除苯酚试验中,温度是主要影响因素,长形水绵去除苯酚试验中,处理时间是主要影响因素;在考虑交互作用下,脆弱刚毛藻和长形水绵对苯酚的去除率分别为88.98%和75.63%,脆弱刚毛藻去除苯酚试验中,温度是主要影响因素,长形水绵去除苯酚试验中,处理时间是主要影响因素;脆弱刚毛藻去除苯酚实验中各因素最优水平组合为温度18℃,处理时间2h,藻重0.5g,长形水绵去除苯酚试验中中各因素最优水平组合为温度8℃,处理时间1h,藻重0.5g。     

Behavioral and Physiological Differences between Two Parapatric Heliconius Species1

The behavior and physiology of two parapatric sibling species, i Heliconius erato cyrbia Godt. and H. himera Hew., were investigated to assess if environmental adaptation enabled stable morphological, genetic, and ecological differences to exist in the face of hybridization. Morning and evening activity, egg production, and larval development time of H. himera and H. erato in insectaries were recorded; individuals were collected in allopatry and in sympatry from a hybrid zone in which the species overlapped. Studies were performed at ambient conditions within the natural range of H. himera. H. himera was considerably more active than H, erato flying earlier in the morning and later in the evening, even when both species were collected in sympatry. Similarly, H. himera laid more eggs, and the hatched larvae developed more rapidly. The results suggest that physiological constraints are an important selective force that may have been important in speciation and counteracts hybridization in the maintenance of the H. himeral H. erato contact zone. Ecological selection, arising from adaptation to low temperatures, may help explain the competitive exclusion of H. erato by H. himera in the drier, cooler montane habitat favored by the latter species.