Mutational analysis of the functional domains of yeast K1 killer toxin.

To determine the functional domains of K1 killer toxin, we analyzed the phenotypes of a set of mutations throughout regions encoding the alpha- and beta-toxin subunits that allow secretion of mutant toxins. A range of techniques have been used to examine the ability of mutant toxins to bind to beta-glucan cell wall receptor and to form lethal ion channels. Our results indicate that both the alpha and beta subunits are involved in beta-glucan receptor binding. Defects in ion channel formation and toxin immunity are confined to the hydrophobic alpha subunit of the toxin.     

A monoclonal antibody that inhibits opioid binding to rat brain membranes

To understand the structure-function relationship and to probe the molecular characteristics of the purified opioid receptor, monoclonal antibodies (mab) were raised against a purified opioid receptor protein. After intensive screening of almost 1500 hybridoma cell lines, only 7 clones were shown to have very high immunoreactivity against the purified receptor. Moreover, out of these 7 clones, only 2, 3B4F11 and 3A27G, were found to inhibit the ligand binding property of the mu-opioid receptor. The mab 3B4F11 was found to inhibit 3H-diprenorphine binding to the purified receptor in a dose dependent manner with a maximal inhibition of 100% achieved with 20 micrograms of the antibody. Likewise, Fab fragments prepared from the mabs 3B4F11 inhibited 3H-diprenorphine binding to P2 membranes in a dose-dependent manner. In addition, it was found that the binding of 3H-DAGO, 3H-DPDPE and 3H-EKC was inhibited with approximately equal potency, suggesting that the Fabs prepared from the mab 3B4F11 interact with all 3 receptor types.     

The interaction of quinone analogues with wild-type and ubiquinone-deficient yeast mitochondria

The interaction of the exogenous quinones, duroquinone (DQ) and the decyl analogue of ubiquinone (DB) with the mitochondrial respiratory chain was studied in both wild-type and a ubiquinone-deficient mutant of yeast. DQ can be reduced directly by NADH dehydrogenase, but cannot be reduced by succinate dehydrogenase in the absence of endogenous ubiquinone. The succinate-driven reduction of DQ can be stimulated by DB in a reaction inhibited 50% by antimycin and 70-80% by the combined use of antimycin and myxothiazol, suggesting that electron transfer occurs via the cytochrome b-c1 complex. Both DQ and DB can effectively mediate the reduction of cytochrome b by the primary dehydrogenases through center o, but their ability to mediate the reduction of cytochrome b through center i is negligible. Two reaction sites for ubiquinol seem to be present at center o: one is independent of endogenous Q6 with a high reaction rate and a high Km; the other is affected by endogenous Q6 and has a low reaction rate and a low Km. By contrast, only one ubiquinol reaction site was observed at center i, where DB appears to compete with endogenous Q6. DB can oxidize most of the pre-reduced cytochrome b, while DQ can oxidize only 50%. On the basis of these data, the possible binding patterns of DB on different Q-reaction sites and the requirement for ubiquinone in the continuous oxidation of DQH are discussed.     

The complete amino-acid sequence of the proteinase inhibitor B from the root of the arrowhead (Sagittaria sagittifolia L.)

After reduction and alkylation of the disulfide bonds of the proteinase inhibitor B from the root of the arrowhead (Sagittaria sagittifolia L.) followed by CNBr cleavage three peptide fragments with 68, 62 and 11 amino-acid residues could be separated on DEAE-Sepharose CL-6B. The peptides or the inhibitor itself were further specifically cleaved either by trypsin or by the mixture of (CH3)2SO/HCl/HBr at the arginyl- and the tryptophyl-peptide bond, respectively. The complete amino-acid sequences of the peptides were determined by manual solid phase DABITC/PITC double coupling micro-method and the primary structure of the arrowhead inhibitor B consisting of 141 amino-acid residues was then elucidated. Twenty pairs of amino-acid residues are repeated in the molecule of this inhibitor, three of these pairs even occur three times. The possible locations of the reactive sites are discussed. On the basis of sequence comparisons between this inhibitor and all other serine proteinase inhibitors the arrowhead inhibitor may belong to a new family.     

甲型流感病毒自然温度敏感株的宿主依赖性

用鸡胚细胞(CEC)和Madin-Darby Canine Kidney(MDCK)两个宿主细胞系统,检查甲型流感病毒各亚型不同时期流行株的温度敏感性状(ts),发现自然界存在许多宿主依赖的温度敏感性突变株(hd-ts)。它们在CEC上是ts,在MDCK上却是ts~ 。检出的hd-ts株中有些曾经过人体接种观察证明为减毒株。这表明在CEC中鉴定的ts表型与对人减毒性状密切相关。     

贵州部分森林群落物种多样性初步研究

 本文根据贵州省178个森林群落样地的数据研究了群落物种的多样性。测定的指标有群 落物种丰富度,群落Simpson多样性指数和群落均匀度。测定结果表明;不同垂直带生物气 候条件下的森林群落有不同的多样性。相同垂直带生物气候条件下,基质生境相同时,不同森林植被亚型的群落的多样性近似,基质生境不同时,群落多样性则不同;同一群落类型的各个样地的多样性也有变化,结构不同的群落个体,其多样性指数不同,演替趋势也不同。乔木第二亚层的多样性普遍地高于乔木第一亚层。同一演替系列中,越接近顶极阶段多样性越高。多样性指数与群落物种丰富度,均匀度呈紧密的正相关,与群落个体总数没有相关。认为多样性测定在比较、说明群落的结构、类型、组织特征、生境、演替等方面有一定意义。     

细胞色素C_3在Rhodopseudomonas caosulata载色体光合磷酸化中的作用

细胞色素C_3不仅能增进除去铁氧还蛋白载色体的循环光合磷酸化活性的恢复,而且也增进以DCPIPH_2为电子供体,维生素K_3、反丁烯二酸分别为电子受体构成的非循环光合磷酸化活性的恢复。 氩气相下,细胞色素C_3促进正常载色体光合磷酸化活性的最适浓度是1.8 μmol,当PMS存在时,这一促进作用随C_3浓度的增加而直线上升,然后呈稳态。 Antimycin A(10~(-7)mol/L)能充分抑制C_3参与的光合磷酸化活性,这一抑制现象在PMS存在时消失。 o~-phenanthroline(1×10~(-5)mol/L.)对C_3参与的光合磷酸化活性亦具抑制作用,并被PMS的添加而消失,当浓度提高时(10~(-3)mol/L),抑制现象不因PMS的存在而消失。 氢气相下的载色体光合磷酸化活性比氩气相下的低,并且随着载色体贮存(-20℃)时间的增长而急剧下降,24h贮存,丧失活性达60％。C_3对氢气相下的光合磷酸化活性具明显的促进作用,而铁氧还蛋白则不能,但两者同时存在时,其磷酸化活性显著提高。     

Ti质粒T区tms基因的分离及其对高等植物分化的影响

根癌农杆菌Ti质粒的T区DNA带有致瘤基因,其基因1和基因2编码生长素吲哚乙酸生物合成途径中的两个酶。以pGV 354(pBR322质粒中插有Ti质粒C 58 T区DNA的HindⅢ15—HindⅢ22大片段)重组质粒出发,我们分离了基因1和基因2,并构建了带有卡那霉素抗性基因的重组质粒pBZ 692,通过基因载体pGV 3850,我们将基因1和基因2引入了高等植物。结果证明基因1和基因2能促使烟草、向日葵、土豆等转化组织分化长根,转化的根在MS_0培养基上能脱分化形成愈伤组织并自主生长,在转化的组织中有转化标记胭脂碱的存在。     

Fluorescence polarization as a tool to study lectin-sugar interaction

The binding ofRicinus communis agglutinin andAbrus agglutinin to 4-methylumbelliferyl β-D-galactopyranoside was studied by equilibrium dialysis, fluo-rescence quenching and fluorescence polarization. The number of binding sites and the association constant value obtained by fluorescence polarization for bothRicinus communis agglutinin andAbrus agglutinin are in close agreement with those obtained by the other methods. This indicates the potential of ligand-fluorescence polarization measurements in the investigation of lectin-sugar interactions.     

Lectins from<Emphasis Type="Italic">Griffonia simplicifolia</Emphasis> seeds

The physical-chemical and carbohydrate binding specificity ofGriffonia simplicifolia I (GS I) isolectins, one of the 4 lectins isolated fromGriffonia simplicifolia seeds, are described.Association constants for the binding of methyl α- and β-D-galactopyranoside and methyl 2-acetamido-2-deoxy-α-D-galactopyranoside to the A₄, A₂ B₂ and B₄ isolectins are reported.Precipitation reactions of theGriffonia simplicifolia isolectins with guaran and type B blood group substance are described.The hypothesis that subunit B is a precursor of subunit A, a process involving proteolytic cleavage of the B subunit, was tested by conducting structural studies on the 2 subunits. The results indicated that the A and B subunits are probably products of 2 separate but closely related, possibly contiguous genes.