Catalytic properties of chloroplast F1-ATPase modified at catalytic or noncatalytic sites by 2-azido adenine nucleotides

When heat-activated F1-ATPase from chloroplasts was repeatedly exposed to Mg2+ and 2-azido-ATP, followed by separation from medium nucleotides and photolysis, a total of two sites per enzyme, both catalytic and noncatalytic, were labeled. In a coupled assay with pyruvate kinase about half the activity was lost when one site per enzyme was modified. However, increased modification resulted in no further loss of activity. In contrast, methanol-sulfite activation of the enzyme showed a loss of most of the catalytic capacity when one site per enzyme was modified. Predominant labeling of either one catalytic or one noncatalytic site caused a loss of most of the activity in either assay. An indication that the enzyme modified at one site retained some catalytic activity was verified by measurement of the [18O]Pi species formed when [gamma-18O]ATP was hydrolyzed by partially derivatized enzyme. With either catalytic or noncatalytic site modification, the distributions of [18O]Pi species formed showed that the modified enzyme had different catalytic characteristics. An interpretation is that with modification by azido nucleotides at either catalytic or noncatalytic sites, capacity for rapid catalysis is largely lost but the remaining sites retain weak modified catalytic properties.     

Cross-reactive cellular and humoral immunity to carcinogen-induced chicken fibrosarcomas. I. Protective cross-immunity between transplantable tumor lines

Dimethylbenz[a]anthracene (DMBA)-induced transplantable fibrosarcomas in B2 homozygous chickens (SC line) grow progressively in normal chickens, but are rejected by chickens immunized previously with irradiated tumor cells and Corynebacterium parvum. Tumor-immune chickens resist challenge by the immunizing tumor lines as well as by some, but not all, fibrosarcoma lines. The pattern of cross-reactivity between four DMBA-induced transplantable tumor lines was examined in detail. Ability to reject a tumor challenge correlated very well (p less than 0.001) with the presence of delayed-type hypersensitivity (DTH) to that tumor. Immunization with one of two of the DMBA-induced lines tested also caused rejection of transplantable tumors developed from methylcholanthrene-induced and benzo(a)pyrene-induced primary fibrosarcomas. Although immunization with tumor caused DTH to chicken embryo fibroblasts (CEF), immunization with CEF failed to cause protective immunity or DTH to tumors. Presence of protective immunity, where tested, also correlated with the ability of spleen cells from immune donors to inhibit tumor growth in Winn tests. Humoral immunity exhibited even greater cross-reactivity than did cellular immunity. Distinct patterns of cross-reactivity were nevertheless observed with respect to the serum antibodies as detected in ELISA. Two of these patterns were also observed in several sera from primary tumor-bearing chickens, both including reactivity with CEF. Such reactivity was absent from normal chicken sera.     

Regulation of methyl group metabolism in lactating ewes

The effect of lactation on a number of enzymes involved in transmethylation reactions and the secretion of major methyl compounds into milk have been examined in sheep. The activities of hepatic phospholipid methyltransferase and 5-methyltetrahydrofolate-homocysteine methyltransferase were significantly higher in lactating ewes, compared with those in non-lactating ewes, while the activity of both hepatic and pancreatic glycine methyltransferase was significantly lower in the lactating state. No differences were observed in the activities of hepatic guanidoacetate methyltransferase, betaine-homocysteine methyltransferase and cystathionine beta-synthase on lactation. These results suggest that the extra demand for methyl groups for the secretion of methyl compounds in the milk is facilitated by enhancing the rate of de novo methyl group synthesis and lowering the rate of physiologically nonessential methylation.     

Expression of the adrenergic phenotype by dorsal root ganglion cells of the quail in culture in vitro

From the results of previous studies, we have suggested that "autonomic" cell precursors exist in latent form in sensory ganglia of avian embryos. The potentialities can be expressed when the ganglia are transplanted into a young embryo host. In the present study, we have observed a similar transformation in cultures of dissociated dorsal root ganglia taken from quail embryos of 7-15 days of incubation. From the 4th day of culture onward, numerous adrenergic cells appear. They display tyrosine hydroxylase immunoreactivity, synthesise and store catecholamines and generally differ in size and shape from primary sensory neurons. They and/or their precursors can actively proliferate in culture. The differentiation of these catecholaminergic cells, which can not be detected in quail dorsal root ganglia during normal development in vivo, is dependent on one or more factors present in 9-day chick embryo extract.     

微核形成与细胞周期关系的初步研究 Ⅲ.丝裂霉素c诱发人淋巴细胞染色体畸变与不同周期阶段的微核形成

为了研究染色体畸变与微核形成的关系,本实验用不同浓度的丝裂霉素C(MMC,0.025—0.4μg/ml),处理人外周血淋巴细胞,观察中期染色体畸变与不同细胞周期形成的微核间的关系。获得如下主要结果:(1)MMC诱发的染色体畸变细胞率(ACF),未经培养的G_0期淋巴细胞的微核细胞率(NC-MNCF)以及培养的淋巴细胞的微核细胞率(C-MNCF),在一定剂量范围内均呈剂量依赖性增加,并可用幂回归方程描述;(2)微核形成与染色体畸变全然无关的NC-MNCF,和C-MNCF一样,与ACF呈良好的正相关;(3)用胞质分裂阻滞(CB)法,检测MMC诱发的CB-MNCF,较C-MNCF无显著提高,MNCF/ACF的比值较小,并随着MMC剂量增加从0.15左右降到0.03。所有上述结果表明,不能简单理解微核形成与染色体畸变间的关系,在分裂的细胞群体中,中期染色体畸变可能仅是微核形成的一种来源。     

A covalently constrained congener of the Saccharomyces cerevisiae tridecapeptide mating pheromone is an agonist

An analog of alpha-factor, the Saccharomyces cerevisiae tridecapeptide mating pheromone (Trp-His-Trp-Leu-Gln-Leu-Lys-Pro-Gly-Gln-Pro-Met-Tyr), in which the side chains of Lys7 and Gln10 were covalently linked, was synthesized using solid phase methodologies. The yield of the purified cyclic analog cyclo7,10[Nle12]alpha-factor was 30%, and its structure was verified by amino acid analysis, peptide sequencing, fast atom bombardment-mass spectrometry, and proton nuclear magnetic resonance spectroscopy. Cyclo7,10[Nle12]alpha-factor caused growth arrest and morphological alterations in S. cerevisiae MATa cells qualitatively identical to those induced by linear pheromone and was one-fourth to one-twentieth as active as the linear alpha-factor depending upon the S. cerevisiae strain tested. Consistent with the relative activities of the linear and cyclic peptides, binding competition studies indicated that cyclo7,10[Nle12]alpha-factor had approximately 20-40-fold less affinity for the alpha-factor receptor. Hydrolysis of the cyclic peptide by the target cells did not lead to opening of the ring and was less rapid than that of linear alpha-factor. The alpha-factor antagonist des-Trp1-[Ala3,Nle12]alpha-factor reversed the activity of the cyclic analog, and cyclo7,10[Nle12]alpha-factor was not active at the restrictive temperature in a temperature-sensitive receptor mutant. These results support the conclusion that the cyclic alpha-factor occupies the same binding site within the receptor as is occupied by the natural pheromone. The cyclic alpha-factor represents a rare example of an agonist among covalently constrained congeners of small linear peptide messengers.     

Phosphorylation and nuclear localization of the varicella-zoster virus gene 63 protein.

The protein encoded by varicella-zoster virus open reading frame 63 and carboxy-terminal deletions of the same were expressed either as fusion proteins at the carboxy terminus of the maltose-binding protein in Escherichia coli or independently in transfected mammalian cells. The truncations contained amino acids 1 to 142 (63 delta N) or 1 to 210 (63 delta K) of the complete 278-amino-acid primary sequence. Recombinant casein kinase II phosphorylated the 63F and 63 delta KF fusion proteins in vitro but did not phosphorylate the 63 delta NF fusion protein, implying that phosphorylation occurred between amino acids 142 and 210. Immunoprecipitation of 35S- or 32P-labelled extracts of cells transfected with plasmids expressing 63, 63 delta N, or 63 delta K also indicated that in situ phosphorylation most likely occurred between amino acids 142 and 210. These combined results suggest that casein kinase II plays a significant role in the phosphorylation of the varicella-zoster virus 63 protein. Indirect immunofluorescence of transfected cells indicated nuclear localization of the 63 protein and cytoplasmic localization of 63 delta K and 63 delta N, implying a requirement for sequences between amino acids 210 and 278 for efficient nuclear localization.     

几种昆虫生长调节剂对家白蚁的毒效试验

在室内条件下测定了卡死克、抑太保、灭幼豚3号、爱力螨克和扑虱灵五种昆虫生长调节剂对家白蚁的毒杀效果。初步筛选结果表明：卡死克、抑太保和爱力螨克对家白蚁的毒杀效果均较好,家白蚁对爱力螨克尤其敏感。2．30pm。yL爱力螨克、327．36pm0VL、卡死克和369．80V＊wL抑太保处理白蚁5～6天后,其死亡率可达100％。忌避性试验表明：卡死克、抑太保和爱力螨克对家白蚁均无明显的驱避作用。