The enzymic nature of antibody catalysis: development of multistep kinetic processing

Detailed kinetic investigations of a catalytic antibody that promotes the hydrolyses of an anilide and phenyl ester show that this catalyst uses a multistep kinetic sequence resembling that found in serine proteases to hydrolyze its substrates, although antibody was elicited to a single transition-state analog. Like the serine proteases the antibody catalyzes the hydrolysis reactions through a putative covalent intermediate, but unlike the enzymes it may use hydroxide ion to cleave the intermediates. Nevertheless, the antibody is a potent catalyst with turnover at higher pH values rivaling that of chymotrypsin. This analysis also reveals that turnover by the antibody is ultimately limited by product desorption, suggesting that improvements in catalytic efficiency may be achieved by judicious changes in the structure of the substrate, so that it is not superimposable on that of the eliciting hapten.     

Tumor etiology and chromosome pattern

Fibrosarcomas induced in Chinese hamsters and rats by Rous sarcomla virus and 7,12-dimethylbenz(a)anthracene are associated with nonrandom chromosome variation. Although histologically indistinguishable, the tumors induced by the virus or chemical in each host species are characterized by completely different karyotypic patterns.     

Epoxides of carcinogenic polycyclic hydrocarbons are frameshift mutagens

K-region epoxides of the carcinogens benz[a] anthracene, dibenz[a,h]anthracene, and 7-methylbenz[a] anthracene are mutagenic in strains of Salmonella typhimurium designed to detect frameshift mutagens. Parent hydrocarbons, K-region diols and phenols and some other epoxides are inactive as mutagens in these tests. Polycyclic hydrocarbon epoxides, and other presumed proximal carcinogens, are discussed as examples of intercalating agents with reactive side chains. It has been shown previously that intercalating agents with reactive side chains are potent frameshift mutagens.     

Role of the protein moiety of ribonuclease P, a ribonucleoprotein enzyme

The Bacillus subtilis ribonuclease P consists of a protein and an RNA. At high ionic strength the reaction is protein-independent; the RNA alone is capable of cleaving precursor transfer RNA, but the turnover is slow. Kinetic analyses show that high salt concentrations facilitate substrate binding in the absence of the protein, probably by decreasing the repulsion between the polyanionic enzyme and substrate RNAs, and also slow product release and enzyme turnover. It is proposed that the ribonuclease P protein, which is small and basic, provides a local pool of counter-ions that facilitates substrate binding without interfering with rapid product release.     

5'-腺苷磷酰硫酸激酶及其R68K突变体对5'-腺苷一磷酸3'羟基的磷酸化

硫作为生命活动的必需元素,主要以-2价和+6价发挥生物学功能。硫的同化代谢包括胞内活化、转移以及还原等反应。其活化是同化代谢的关键反应,包括ATP硫酸化酶(ATP sulfurylase,ATPS)催化硫酸盐与ATP反应生成腺苷-5'-磷酰硫酸(adenosine 5'-phosphosulfate,APS)和焦磷酸(pyrophosphate,PPi)以及腺苷-5'-磷酰硫酸激酶(adenosine 5'-phosphosulfate kinase,APSK)催化APS 3'羟基磷酸化生成3'-磷酸腺苷-5'-磷酰硫酸(3'-phosphoadenosine 5'-phosphosulfate,PAPS),APSK催化APS磷酸机理已经较为清楚。利用APSK对AMP的磷酸化进行了初步分析,发现AMP可作为APSK的底物,反应生成3,5'二磷酸腺苷(3'-phosphoadenosine 5'-phosphate,PAP);对APSK的三维结构进行分析发现,R68同时和APS的磷酸根和硫酸根形成氢键,稳定APS的结合,而K侧链基团比R短2.4魡,R68K突变将导致K不能和距离较远的硫酸根离子相互作用,从而减弱APS的亲和力,而增加与磷酸根离子的相互作用,可能提高AMP的亲和力。研究结果表明,R68K突变体的最适底物变为AMP,KmAMP是对照的0.2倍,而催化效率是对照的5倍。以R68K为偶联酶成功测定了具有较低KmPAP的酵母3,5二磷酸核苷酸酶(3',5'-bisphosphate nucleotidase,YND)动力学常数,为分析测定AMP底物的酶活提供了工具。     

Regulation of Ras-GAP and the neurofibromatosis-1 gene product by eicosanoids

Ras-GAP (GTPase activating protein) is a regulatory protein that stimulates the intrinsic guanosine triphosphatase (GTPase) activity of the proto-oncogene product p21ras. A domain of the neurofibromatosis gene product (NF1) that has sequence similarity to the catalytic domain of Ras-GAP and to yeast IRA gene products also has a specific stimulatory activity toward p21ras GTPase. Arachidonic acid and phosphatidic acid inactivate GAP, but no agents have been identified that stimulate GAP and thereby switch p21ras off. With the use of recombinant Ha-c-Ras and Ras-GAP, NF1, and GAP catalytic domains, it was found that prostaglandins PGF2 alpha and PGA2 stimulated Ras-GAP and that prostacyclin PGI2 inhibited Ras-GAP. The stimulatory effect of PGF2 alpha was saturable and structure-specific and competed with the inhibitory effect of arachidonic acid. Arachidonic acid also inhibited the catalytic activity of NF1, but prostaglandins were not stimulatory. These results suggest a mechanism for the allosteric control of Ras function through the modulation of arachidonate metabolism.     

降解蒽遗传重组菌在蒽污染环境中的降解和存活能力研究

为了提高外源蒽降解菌An降解蒽的能力和存活能力，采用原生质体电融合技术，在蒽降解菌An和土生优势菌Tu之间实行了原生质体电融合，并从再生的融合子F中筛选到具有特定性状的后代菌株。把融合子接种于蒽污染的土壤与水环境中，研究其降解蒽的能力与存活能力。结果表明，在蒽污染的土壤中，40d时，融合子F与亲本An对蒽的降解率分别为70．7％、55．5％，融合子比出发菌株高15．2％；100h内融合子F和亲本An对水环境中蒽的降解转化率分别为75％、67．2％。说明在蒽污染的土壤和水环境中．在初始接种量相同的情况下，融合子的存活率明显高于出发菌株。     

Enzyme dynamics during catalysis

Internal protein dynamics are intimately connected to enzymatic catalysis. However, enzyme motions linked to substrate turnover remain largely unknown. We have studied dynamics of an enzyme during catalysis at atomic resolution using nuclear magnetic resonance relaxation methods. During catalytic action of the enzyme cyclophilin A, we detect conformational fluctuations of the active site that occur on a time scale of hundreds of microseconds. The rates of conformational dynamics of the enzyme strongly correlate with the microscopic rates of substrate turnover. The present results, together with available structural data, allow a prediction of the reaction trajectory.     

Accelerated intestinal epithelial cell turnover: a new mechanism of parasite expulsion

The functional integrity of the intestinal epithelial barrier forms a major defense against invading pathogens, including gastrointestinal-dwelling nematodes, which are ubiquitous in their distribution worldwide. Here, we show that an increase in the rate of epithelial cell turnover in the large intestine acts like an "epithelial escalator" to expel Trichuris and that the rate of epithelial cell movement is under immune control by the cytokine interleukin-13 and the chemokine CXCL10. This host protective mechanism against intestinal pathogens has implications for our wider understanding of the multifunctional role played by intestinal epithelium in mucosal defense.     

Catalysis: new perspectives from surface science

One-sixth of the value of all goods manufactured in the United States involves catalytic processes. However, in spite of this dramatic economic impact, little is known about this broad subject at the molecular level. In the last two decades a variety of techniques have been developed for studying at the atomic level the structure, composition, and chemical bonding at surfaces. These techniques have been used to study adsorption and reaction on metal single crystals in an ultrahigh vacuum environment or to analyze catalysts before and after reaction. An important new development has been the coupling of an apparatus for the measurement of reaction kinetics at elevated pressures with an ultrahigh vacuum system for surface analysis. This approach has demonstrated that metal single crystals can be used to successfully model many important catalytic reactions and has established a direct link between the results of ultrahigh vacuum surface measurements and the chemistry that occurs under typical catalytic-processing conditions.     

Quantum impurity in a nearly critical two-dimensional antiferromagnet

The spin dynamics of an arbitrary localized impurity in an insulating two-dimensional antiferromagnet, across the host transition from a paramagnet with a spin gap to a Neel state, is described. The impurity spin susceptibility has a Curie-like divergence at the quantum-critical coupling, but with a universal effective spin that is neither an integer nor a half-odd integer. In the Neel state, the transverse impurity susceptibility is a universal number divided by the host spin stiffness (which determines the energy cost to slow twists in the orientation of the Neel order). These and numerous other results for the thermodynamics, Knight shift, and magnon damping have important applications in experiments on layered transition metal oxides.     

催化铁-生物耦合处理草甘膦废水生物出水的可行性研究

[目的]针对草甘膦生产过程中排出的废水有机物和有机磷浓度高、可生化性差,传统工艺难以达标的问题,进行催化铁还原技术的可行性研究,为现有处理工艺的改造提供参考。[方法]通过摇瓶试验和SBR持续试验,对催化铁还原技术和微生物处理过程耦合处理现有草甘膦废水生物出水COD和磷的去除效果进行研究。[结果]将催化铁还原技术与微生物处理过程耦合能够提高草甘膦农药废水的COD和TP去除效果;耦合的SBR系统在微氧条件下对原厌氧出水的COD和TP去除率分别稳定在80.0%和50.0%以上,正磷酸盐浓度在20.00 mg/L以下。[结论]催化铁-生物耦合为草甘膦废水的处理和改造提供了一种较好的处理方法。     

Autoradiographic imaging of phosphoinositide turnover in the brain

With [3H]cytidine as a precursor, phosphoinositide turnover can be localized in brain slices by selective autoradiography of the product [3H]cytidine diphosphate diacylglycerol, which is membrane-bound. In the cerebellum, glutamatergic stimulation elicits an increase of phosphoinositide turnover only in Purkinje cells and the molecular layer. In the hippocampus, both glutamatergic and muscarinic cholinergic stimulation increase phosphoinositide turnover, but with distinct localizations. Cholinergic stimulation affects CA1, CA3, CA4, and subiculum, whereas glutamatergic effects are restricted to the subiculum and CA3. Imaging phosphoinositide turnover in brain slices, which are amenable to electrophysiologic studies, will permit a dynamic localized analysis of regulation of this second messenger in response to synaptic stimulation of specific neuronal pathways.     

植物纤维/塑料复合材作地板基材的研究

利用植物纤维与塑料加工生产地板材料的试验表明,利用纤维/聚丙稀复合的产品有较好的性能,可以满足合格品的质量要求,在添加偶联剂的条件下,可满足优等品的质量要求 而其余2种塑料与纤维的复合在添加偶联剂的条件下可超过合格品的质量要求 偶联剂的存在,改善了纤维与塑料的界面状态,提高了复合材料的玻璃转化温度,从而提高了复合材料的性能     

Chimeras of two isoprenoid synthases catalyze all four coupling reactions in isoprenoid biosynthesis

The carbon skeletons of over 55,000 naturally occurring isoprenoid compounds are constructed from four fundamental coupling reactions: chain elongation, cyclopropanation, branching, and cyclobutanation. Enzymes that catalyze chain elongation and cyclopropanation are well studied, whereas those that catalyze branching and cyclobutanation are unknown. We have catalyzed the four reactions with chimeric proteins generated by replacing segments of a chain-elongation enzyme with corresponding sequences from a cyclopropanation enzyme. Stereochemical and mechanistic considerations suggest that the four coupling enzymes could have evolved from a common ancestor through relatively small changes in the catalytic site.     

Transition state stabilization by a catalytic RNA

The hairpin ribozyme catalyzes sequence-specific cleavage of RNA through transesterification of the scissile phosphate. Vanadate has previously been used as a transition state mimic of protein enzymes that catalyze the same reaction. Comparison of the 2.2 angstrom resolution structure of a vanadate-hairpin ribozyme complex with structures of precursor and product complexes reveals a rigid active site that makes more hydrogen bonds to the transition state than to the precursor or product. Because of the paucity of RNA functional groups capable of general acid-base or electrostatic catalysis, transition state stabilization is likely to be an important catalytic strategy for ribozymes.     

Mammary cancer induction by 7,12-dimethylbenz(a)anthracene: relation to age

Mammary glands were transplanted from donors treated with 7,12-dimethylbenz(a)anthracene to untreated isologous recipients. Incidence of mammary tumors in mammary grafts from 56-day-old donors was significantly higher than that in grafts from 120-day-old donors, regardless of the age of the recipient hosts. Whent mammary glands were transplanted from untreated donors to isologous recipients that subsequently received 7,12-dimethyl-benz(a)anthracene, a similar difference in tumor incidence in the grafts was observed. In contrast to mammary glands of older females, mammary glands of young adult female rats are highly vulnerable to the carcinogenic effect of 7,12-dimethylbenz(a)anthracene.     

江西五倍子蚜虫冬寄主的初步研究

五倍子为东南亚特产，具有广泛的应用领域，我国尤以药用名。本报道江西产五倍子种类的同时，记述了江西五倍子蚜虫冬寄主的种类及分布，为本省五倍子生产，提供冬寄主环节的初步依据。     

Efficient activation of aromatic C-H bonds for addition to C-C multiple bonds

Efficient electrophilic metalation of aromatic C-H bonds leading to new C-C bond formation through regio- and stereoselective addition to alkynes and alkenes has been realized by a catalytic amount (0.02 to 5 mole percent) of palladium(II) or platinum(II) compounds in a mixed solvent containing trifluoroacetic acid at room temperature. Various arenes undergo unexpected selective trans hydroarylation to terminal or internal C&cjs0812;C bonds inter- and intramolecularly with high efficiency (up to a turnover number of 4500 for palladium), especially for electron-rich arenes, giving thermodynamically unfavorable cis-alkenes, and the oxygen- and nitrogen-containing heterocycles. The simplicity, generality, and efficiency of this process should be very attractive to the possible industrial application for the functionalization of arenes.