The reaction of cytochromes c and c2 with the Rhodospirillum rubrum reaction center involves the heme crevice domain

In order to define the interaction domain on Rhodospirillum rubrum cytochrome c2 for the photosynthetic reaction center, positively charged lysine amino groups on cytochrome c2 were modified to form negatively charged carboxydinitrophenyl lysines. The reaction mixture was separated into six different fractions by ion exchange chromatography on carboxymethylcellulose and sulfopropyl-Sepharose. Peptide mapping studies indicated that fraction A consisted of a mixture of singly labeled derivatives modified at lysines 58, 81, and 109 on the back of cytochrome c2. Fractions C1, C2, C3, and C4 were found to be mixtures of singly labeled derivatives modified at lysines 9, 13, 75, 86, and 88 on the front of cytochrome c2 surrounding the heme crevice. The photooxidation of the carboxydinitrophenyl-cytochrome c2 derivatives by reaction centers purified from R. rubrum was measured following excitation with a laser pulse. The second-order rate constant of fraction A modified at backside lysines was found to be 2.3 X 10(7) M-1 s-1, nearly the same as that of native cytochrome c2, 2.6 X 10(7) M-1 s-1. However, the rate constants of fractions C1-C4 were found to be 6 to 12-fold smaller than that of native cytochrome c2. These results indicate that lysines surrounding the heme crevice of cytochrome c2 are involved in electrostatic interactions with carboxylate groups at the binding site of the reaction center. The reaction rates of horse heart cytochrome c derivatives modified at single lysine amino groups with trifluoroacetyl or trifluoromethylphenylcarbamoyl were also measured. Modification of lysines 8, 13, 25, 27, 72, 79, or 87 surrounding the heme crevice was found to significantly lower the rate of reaction, while modification of lysines in other regions had no effect. This indicates that the reaction of horse heart cytochrome c with the reaction center also involves the heme crevice domain.     

Linkage map of seven polymorphic markers on rat Chromosome 18

A genetic linkage map of seven polymorphic markers was created with F₂ intercross progeny of F344/N and LEW/N rats and assigned to rat Chromosome (Chr) 18. Five of the markers described were defined by simple sequence length polymorphisms (SSLPs) associated with five genes: transthyretin (TTR), trypsin inhibitor-like protein (TILP), 2 adrenergic receptor (ADRB2), olfactory neuron-specific G protein (OLF), and gap junction protein (GJA1). One marker was defined by a restriction fragment length polymorphism (RFLP) detected with a probe for the human colony stimulating factor 1 receptor (CSF1R) gene. The D18N1R locus was defined by an anonymous DNA fragment amplified by the randomly amplified polymorphic DNA (RAPD) technique with a single short primer. These seven DNA loci formed a single genetic linkage group 30.4 cM in length with the following order: TTR-6.8 cM-D18N1R-9.1 cM-TILP-4.3 cM-CSF1R-0 cM-ADRB2-10.2 cM-OLF-0 cM-GJA1. The five SSLP markers were highly polymorphic. In a total of 13 inbred rat strains analyzed (F344/ N, LEW/N, LOU/MN, WBB1/N, WBB2/N, MR/N, MNR/N, ACI/N, SHR/N, WKY/N, BN/SsN, BUF/N, and LER/N), three to six alleles were detected for each marker. Remarkable linkage conservation was detected between the region of rat Chr 18 mapped and a region of mouse Chr 18. However, genes associated with these markers have been mapped to three different human chromosomes (Chrs 5, 6, and 18). The markers described here should be useful for genetic mapping studies and genetic monitoring of inbred rat strains.     

视黄酸和亚硒酸钠对肝癌细胞某些表型逆转的加和作用

 人肝癌细胞株SMMC-7721经1μmol/L视黄酸和或2.5μmol/L亚硒酸钠处理后,膜上纤维连接蛋白沉着量逐日上升,且较相应天数的对照组细胞增加,而甲胎蛋白分泌量和~3H-TdR参入率被明显抑制。视黄酸和亚硒酸钠同时处理的联合组作用强度接近于两者单独使用时作用强度的加和。对以上结果和视黄酸及亚硒酸钠使肝癌细胞接触抑制恢复及表型逆转的关系作了讨论。     

棉铃虫的取食营养特点与棉花抗虫素分布的关系

通过研究棉花8个部位对棉铃虫Helicoverpa armzgera的营养效果和其所含次生物质类萜烯和单宁的浓度及在组织中的分布,揭示了棉铃虫取食营养特点与棉花次生化学的关系。棉铃虫取食顶尖、转移蛀食蕾铃的习性,与有关器官或组织对幼虫的营养效果密切相关,而营养效果主要取决于类萜烯和单宁的含量。棉花顶尖嫩叶中单宁浓度随着棉花的生长发育呈升高的趋势;类萜烯浓度在第四真叶期、第六真叶期和现蕾初期之间出现一个明显的底谷,而此期幼虫主要为害顶尖。蕾铃外层的苞叶、花萼、花瓣和铃皮,因次生物质含量高,不利于幼虫生长,相反内部的花粉、子房和铃心,次生物质含量低,营养效果好,顶尖嫩叶则介于其间。类萜烯存在于组织的色素腺内,分布集中;单宁则散布于组织中,偏多分布于组织外层。结果证实,昆虫在寄主植物上的取食方式是昆虫对寄主体内变动的次生化学的一种适应,它使昆虫付出尽少代价获得最适营养效果。     

成人腹泻轮状病毒的提纯及其高价免疫血清的制备

本文采用交叉免疫电泳技术,首先提纯了成人腹泻轮状病毒(Adult DiarrhoeaRotavirus简称:ADRV),并经电镜确认,用作免疫抗原,首次制备了高价兔与豚鼠抗ADRV血清和小鼠抗ADRV腹水,其特异性经交叉免疫电泳和对流免疫电泳鉴定,其效价经酶联免疫吸附试验(ELISA)检测均在1:2,000以上。     

Arbuscular mycorrhizal fungus enhances P acquisition of wheat (Triticum aestivum L.) in a sandy loam soil with long-term inorganic fertilization regime

The P efficiency, crop yield, and response of wheat to arbuscular mycorrhizal fungus (AMF) Glomus caledonium were tested in an experimental field with long-term (19 years) fertilizer management. The experiment included five fertilizer treatments: organic amendment (OA), half organic amendment plus half mineral fertilizer (1/2 OM), mineral fertilizer NPK, mineral fertilizer NK, and the control (without fertilization). AMF inoculation responsiveness (MIR) of wheat plants at acquiring P were estimated by comparing plants grown in unsterilized soil inoculated with G. caledonium and in untreated soil containing indigenous AMF. Without AMF inoculation, higher crop yields but lower colonization rates were observed in the NPK and two OA-inputted treatments, and NPK had significantly (P < 0.05) lower impacts on organic C and available P in soils and thereby P acquisition of wheat plants compared with OA and 1/2 OM. G. caledonium inoculation significantly (P < 0.05) increased colonization rates with the NPK and two P-deficient treatments but significantly (P < 0.05) increased vegetative biomass, crop yield, and P acquisition of wheat as well as soil alkaline phosphatase (ALP) activity, only with the NPK treatment. This gave an MIR of ca. 45% on total P acquisition of wheat plants. There were no other remarkable MIRs. It suggested that the MIR is determined by soil available P status, and rational combination of AMF with chemical NPK fertilizer can compensate for organic amendments by improving P-acquisition efficiency in arable soils.     

波罗的海希瓦氏菌AI-2/LuxS对生物被膜和致腐的调控

【目的】波罗的海希瓦氏菌(Shewanella baltica)是冷藏海产鱼类的特定腐败菌。研究群体感应信号AI-2/Lux S对鱼源S.baltica生物被膜和致腐的调控作用。【方法】扩增SB11分离株的lux S基因,用自杀性质粒构建lux S基因缺失株,通过结晶紫染色、珠涡流法、显微镜观察和HPLC,比较分析野生株与缺失株△lux S在4°C和28°C下生物被膜形成、粘附能力、泳动性和致腐产物的差异。【结果】S.baltica SB11中扩增获得lux S基因,生物信息学分析显示Lux S蛋白由169个氨基酸构成,含有保守的His-Thr-Leu-Glu-His(HTLEH)模体和关键氨基酸位点,蛋白三维空间结构与其他细菌相似。与野生株相比,△lux S缺失株上清荧光信号散失,但不影响生长,生物被膜形成期和成熟期的含量显著低于野生株,在4°C培养96 h和28°C培养24 h被膜分别减少20.1%和27.9%。缺失株在不锈钢片的粘附能力明显减弱,其中在4°C培养72 h和28°C培养24 h后的粘附量比野生株分别减少6.48%和6.57%。荧光显微镜观察发现,野生株能快速粘附于玻璃片,聚集形成大量生物被膜,而△lux S仅形成平坦稀疏的被膜,粘附细菌降低,CLSM证实野生株和△lux S的成熟被膜厚度分别为68.95μm和36.44μm。并且,△lux S株在4°C和28°C下泳动性均显著强于野生株。然而,野生株和△lux S株三甲胺和腐胺积累无差异。【结论】鱼源S.baltica中Lux S蛋白保守,AI-2/Lux S参与被膜、粘附能力及泳动性等多种生物被膜形成相关的调控作用,然而不是该菌致腐能力的功能性群体感应信号。     

Novel aminopeptidase N inhibitors derived from antineoplaston AS2–5 (Part I)

Overexpression of zinc-dependent metalloproteinase, aminopeptidase N (APN/CD13), is considered to be involved in the process of tumor invasion and metastasis. Herein we describe the synthesis and in vitro enzymatic inhibition assay of antineoplaston AS2–5 scaffold peptidomimetic compounds. The results demonstrated that most of these l-iso-glutamine derivatives displayed selective inhibitory activity against APN as compared with MMP-2, with IC₅₀ values in the micromole range. The structure–activity relationships were also briefly discussed.