A Ca2+-Dependent Protein Kinase Activity Associated with Serotonin Binding Protein

The endogenous phosphorylation of serotonin binding protein (SBP), a soluble protein found in central and peripheral serotonergic neurons, inhibits the binding of 5-hydroxytryptamine (5-HT, serotonin). A protein kinase activity that copurifies with SBP (SBP-kinase) was partially characterized and compared with calcium/calmodulin-dependent protein kinase II (CAM-PK II). SBP itself is not the enzyme since heating destroyed the protein kinase activity without affecting the capacity of the protein to bind [3H]5-HT. SBP-kinase and CAM-PK II kinase shared the following characteristics: (1) size of the subunits; (2) autophosphorylation in a Ca2+-dependent manner; and (3) affinity for Ca2+. In addition, both forms of protein kinase phosphorylated microtubule-associated proteins well and did not phosphorylate myosin, phosphorylase b, and casein. Phorbol esters or diacylglycerol had no effect on either of the protein kinases. However, substantial differences between SBP-kinase and CAM-PK II were observed: (1) CAM enhanced CAM-PK II activity, but had no effect on SBP-kinase; (2) synapsin I was an excellent substrate for CAM-PK II, but not for SBP-kinase; (3) 5-HT inhibited both the autophosphorylation of SBP-kinase and the phosphorylation of SBP, but had no effect on CAM-PK II. These data indicate that SBP-kinase is different from CAM-PK II. Phosphopeptide maps of SBP and SBP-kinase generated by digestion with S. aureus V8 protease are consistent with the conclusion that these proteins are distinct molecular entities. It is suggested that phosphorylation of SBP may regulate the transport of 5-HT within neurons.     

Use of synthetic DNA in expression of foreign proteins

Synthetic DNAs and oligonucleotides, which can be prepared conveniently by combining chemical synthesis and enzymatic methods, have been used extensively in recombinant DNA research. Examples include total gene synthesis, probes for the isolation of specific genes from cDNA or genomic libraries, linkers containing specific restriction sites for cloning, primers for DNA and RNA sequencing, and primers for the construction of specific mutations (either deletion, insertion or point mutations) by oligonucleotide-directed site-specific mutagenesis.This article reviews recent advances in the chemical and enzymatic synthesis of oligo- and polynucleotides and the application of synthetic DNA to the expression of foreign proteins. The synthesis of genes, including structural genes and regulatory genes are reviewed. Oligonucleotide-directed site-specific mutagenesis and use of synthetic DNA to optimize foreign protein expression are also discussed.     

Hydrodynamic and mechanical aspects of endothelial permeability

The passive transport of water through the endothelial cell layer junctions is considered from the standpoint of hydrodynamic theories based on ultrastructural information. The local geometry of tight junctions based on molecular level forces and elastic membrane properties has been modeled and leads to estimates of the hydraulic resistance of the clefts. It is shown that the large resistance measured experimentally can be accounted for in this model. The transport of large macromolecules via vesicles which diffuse across the endothelial cell has been developed, but recent experimental data do not appear to support this mechanism as a primary pathway. Fused vesicles forming an open channel appear to be rare. Leaky junctions, such as around dying endothelial cells or produced by cytoskeletal changes within the cells, may be important in control of endothelial permeability. Another kind of model is a fiber matrix model of the endocapillary layer, extending into the intercellular clefts which can also account for the molecular seiving properties of the endothelial cell layer but may produce a large resistance to water flux.     

Purification of human C3b inactivator by monoclonal-antibody affinity chromatography

Monoclonal antibody has been obtained to the human complement control protein C3b inactivator after immunization of mice with the enzyme prepared by conventional methods. Antibody from ascitic fluid was purified and coupled to Sepharose-CL-4B to give a specific affinity column, which was used to isolate C3b inactivator from human serum in 70% yield. The product was characterized by size, chain structure, amino acid analysis and proteolytic activity.     

Iodohexestrols. II. Characterization of the binding and estrogenic activity of iodinated hexestrol derivatives, in vitro and in vivo.

The affinity of ortho-iodinated hexestrols for the estrogen binding protein from rat uterus, determined by competitive binding assay, decreases with progressive iodine substitution; 3-iodohexestrol (I-Hex) has a binding affinity 42% that of estradiol. Analysis of [3-H]-I-Hex binding in rat uterine cytosol by sucrose density gradient centrifugation shows both an estrogen-specific binding component (8 S) and a more abundant component (4 S) that is not estrogen specific. Scatchard analysis indicates that this latter binding is of high affinity (Kd equals to 3.7-8.3 times 10- minus-9 M) but is not uterine specific. Polyacrylamide gel electrophoresis shows that most of the [3-H]-I-Hex binding activity in serum and uterine cytosol is distinct from and anodic to the principal protein component (albumin), and that is comigrates with [14-C]thyroxine binding activity. In in vitro incubation of rat uteri, I-Hex can block the specific uptake of [3-H]estradiol into the nuclear fraction; it itself causes a translocation of estrogen-specific binding capacity (as measured by exchange) from cytoplasm to nuclei, and can induce the synthesis of an estrogen-specific uterine protein, all under conditions where it is not metabolically deiodinated to hexestrol. The uterotrophic activities of the iodohexestrols are in most cases comparable to that expected on the basis of their competitive binding affinities. However, selective, estrogen-specific uptake of [3-H]-I-Hex into rat uterus, either in vitro or in vivo, cannot be demonstrated.     

中国淡水养殖池塘环境生态修复技术研究评述

传统的淡水池塘养殖所面临的环境问题日益严重,池塘环境生态修复技术的研究正蓬勃兴起.以“鱼-菜共生”模式为代表的原位修复技术和以循环水养殖模式为代表的异位修复技术是当前研究的重点.本文从水体富营养化的角度,以池塘氮循环为切入点,评述了这两种养殖池塘环境生态修复模式的优缺点,并对其涉及的面积配置关系进行了探讨.评述结果认为,只能通过“鱼-菜共生”模式降低池塘养殖的产排污系数,但通过推广该模式实现全国池塘养殖生态收支的平衡是困难的.虽然循环水养殖模式无法避免产生额外的经济成本和土地资源,但在局部地区,特别是富营养化严重的区域,其零排放的特点使其推广应用有一定的生态价值.     

八角茴香油的提取及应用

本文综述了八角茴香油的主要成分、提取分离方法及在医药等方面的应用,为八角茴香油的进一步研究及应用提供参考。     

水杨酸对小白菜抗铅胁迫的诱导

0.3 mmol/L铅胁迫下小白菜（Brassica rapa var. chinensis）生长受到显著抑制,0.1 mmol/L水杨酸（salicylic acid）诱导处理能显著缓解0.3 mmol/L铅胁迫对小白菜幼苗生长的抑制;并且水杨酸对铅胁迫下小白菜的生长存在“低促高抑”效应,当水杨酸处理浓度达到1.5 mmol/L时,不但不能缓解铅对小白菜的毒害,反而加重毒害,这可能与高浓度水杨酸与铅形成的复合伤害有关。