Characterization of purified cytochrome b-c1 complex from Rhodopseudomonas sphaeroides R-26

A highly purified cytochrome b-c1 complex from Rhodopseudomonas sphaeroides R-26 was isolated by a procedure involving Triton X-100 solubilization, calcium phosphate column chromatography, and ammonium sulfate fractionation. The purified enzyme complex contains, in nanomoles/mg of protein, cytochrome b, 8.3; cytochrome c1, 8.3; iron-sulfur protein, 15; phospholipids, 182; and ubiquinone, 5. Four major polypeptides with apparent molecular weights of 48,000, 30,000, 24,000, and 12,000 were detected in the sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The Mr = 48,000 and 30,000 proteins are cytochromes b and c1, respectively. The enzyme complex catalyzes electron transfer from ubiquinol to cytochrome c with a specific activity of 12.6 mumol of cytochrome c reduced per min/mg of protein at 23 degrees C. This is lower than that of the mitochondrial enzyme, although both systems have similar essential redox components and a similar Km for ubiquinol. The activity is fully sensitive to antimycin A and 5-n-undecyl-6-hydroxy-4, 7-dioxobenzothiazole. The enzyme complex is stable at neutral pH and at lower temperatures, but became less stable when the incubation temperature was raised. At 37 degrees C, the half-life is 15 min. The enzymatic activity was insensitive to treatment with N',N'-dicyclohexylcarbodiimide. No p-chloromercuriphenylsulfonate-alkylable sulfhydryl groups were detected. The major phospholipids associated with the purified enzyme complex are phosphatidylcholine, phosphatidylethanolamine, and phosphatidylglycerol with molar per cent distributions of 25, 21, and 35, respectively. About 60% of the enzymatic activity was abolished upon treatment with phospholipase A2. The phospholipase A2-inactivated activity can be partially restored by the addition of EDTA followed with phospholipids prepared from either the cytochrome b-c1 complex of the same source or a mixture of phosphatidylglycerol and asolectin.(ABSTRACT TRUNCATED AT 250 WORDS)     

水稻叶片衰老过程中氨肽酶活性的变化

水稻叶片中存在着氨肽酶,其最适反应pH和最适反应温度分别为8.2℃和40℃,酶促反应的产物量在最初30min内与时间呈直线相关。 水稻叶片衰老过程中叶绿素和蛋白质含量下降,而氨肽酶比活上升;用植物激素延缓或促进叶片衰老蛋白质降解的同时也抑制或促进了氨肽酶比活的上升,说明氨肽酶在水稻叶片衰老蛋白质降解过程中起一定的作用。根据水稻叶片衰老过程中大分子化合物和叶片外部形态的变化,可将叶片衰老过程划分为缓衰期、急衰期和竭衰期。     

HBsAg重组DNA转化中国地鼠卵巢细胞二氢叶酸还原酶阴性突变株的细胞染色体变化及致瘤性

整合了含乙型肝炎病毒表面抗原(HBsAg)基因和dhfr基因的CHO-dhfr~-细胞,其染色体的畸变率和畸变类型都比亲代CHO-dhfr~-细胞高。但转化前后两系细胞的重要特性都未发生变异,即两者的染色体总数无差别,都是20条。两系细胞株接种裸鼠,均未发现有致瘤性。     

Comparative properties of hydroquinone and hydroxylamine reduction of the Ca(2+)-stabilized O2-evolving complex of photosystem II: reductant-dependent Mn2+ formation and activity inhibition.

Calcium binding to photosystem II slows NH2OH inhibition of O2 evolution; Mn2+ is retained by the O2-evolving complex [Mei, R., & Yocum, C. F. (1991) Biochemistry 30, 7836-7842]. This Ca(2+)-induced stability has been further characterized using the large reductant hydroquinone. Salt-washed photosystem II membranes reduced by hydroquinone in the presence of Ca2+ retain 80% of steady-state O2 evolution activity and contain about 2 Mn2+/reaction center that can be detected at room temperature by electron paramagnetic resonance. This Mn2+ produces a weak enhancement of H2O proton spin-lattice relaxation rates, cannot be easily extracted by a chelator, and is reincorporated into the O2-evolving complex upon illumination. A comparison of the properties of Ca(2+)-supplemented photosystem II samples reduced by hydroquinone or NH2OH alone or in sequence reveals the presence of a subpopulation of manganese atoms at the active site of H2O oxidation that is not accessible to facile hydroquinone reduction. At least one of these manganese atoms can be readily reduced by NH2OH following a noninhibitory hydroquinone reduction step. Under these conditions, about 3 Mn2+/reaction center are lost and O2 evolution activity is irreversibly inhibited. We interpret the existence of distinct sites of reductant action on manganese as further evidence that the Ca(2+)-binding site in photosystem II participates in regulation of the organization of manganese-binding ligands and the overall structure of the O2-evolving complex.     

Epitopes and hemagglutination binding domain on subgenus B:2 adenovirus fibers.

The adenovirus fiber serves as a ligand between the virus and the host cell receptor and manifests hemagglutination (HA) activity and antigenic domains. We have screened both the antigenic and immunogenic epitopes on the adenovirus fibers of subgenus B:2 by using recombinant fiber proteins (rfibers) expressed in Escherichia coli, synthesized peptides (P1 to P8), and the corresponding antisera. The results indicated that P4 (amino acids [aa] 201 to 220), P5 (aa 231 to 250), and P7 (aa 275 to 295) presented both antigenic and immunogenic epitopes in adenovirus type 11 prototype (Ad11p), Ad34a, and Ad11a fibers. P6 (aa 251 to 270) presented both epitopes in Ad11a fiber but only an antigenic epitope in other fibers. The C-terminal 20 amino acids of the fiber, corresponding to P8, manifested an epitope of low-level immunogenicity. P5, localized at the N-terminal aa 231 to 250, displayed an epitope that reacted with fibers of all the members of subgenus B analyzed. The rfibers of Ad11p and Ad34a displayed HA activity with monkey erythrocytes, though those of Ad11a did not. Mutagenesis of the rfibers revealed that neither the fragment replacements, 11p20211a, llp26011a,and 11a28011p, nor the Ad11p rfiber with the substitutions of Tyr-260-->H (Tyr260H)and Arg279Q displayed HA activity. The Ad11a fiber knob was sensitive to proteolytic digestion, whereas that of Ad11p was resistant. The results demonstrated that the decisive HA binding domain was presented at aa 260 to 280 and was conformation dependent. Nearby amino acids, aa 283 and 284, may also affect the HA function.     

利用植物激素调控嫁接形成的初步研究

利用黄瓜（Ｃｕｃｕｍ ｉｓｓａｔｉｖｕｓ）试管苗进行离体茎段自体嫁接,研究ＩＢＡ 和６－ＢＡ 对嫁接形成的影响时发现：进行离体茎段嫁接时,用试管苗茎段可简化嫁接过程,减少污染。嫁接茎段的颜色变化、不定根发生和愈伤组织形成与激素浓度有关。植物激素通过影响砧木和接穗间维管束桥形成的时间和数目调控嫁接组合的发育。在作者的实验中,最佳的激素条件是：在接穗培养基中加ＩＢＡ １．２ ｍ ｇ／Ｌ,在接穗和砧木培养基中加６－ＢＡ ０．３ ｍ ｇ／Ｌ。     

新疆亚鳞木（比较种）角质层特征

报道了产自江苏宜兴晚泥盆世五通组新疆亚鳞木（比较种）Ｓｕｂｌｅｐｉｄｏｄｅｎｄｒｏｎ ｃｆ． ｘｉｎｊｉａｎｇｅｎｓｅＳｕｎ）的表皮角质层用荧光分析显示的特征．该种茎干表皮角质层覆于叶座及叶座间隔带,其中间隔带角质层厚于叶座．表皮细胞在间隔带与叶座表现特征不同．间隔带中部,表皮细胞呈纵长的多边形,其纵长方向与茎干延伸方向相同,细胞壁略有弯曲．间隔带靠近叶座之表皮细胞,细胞壁直,形状类似于前者;但大小仅为前者的１／２左右,且其纵长方向逐渐向叶座边缘偏转．叶座表皮细胞呈近等多边形,有胞间隙．该种茎表皮无气孔     

A stochastic model for the sigmoidal behaviour of cooperative biological systems

A stochastic model for cooperative transitions in biological systems based on a Markov chain is proposed. This model requires only two parameters, the mean probability, p, and the coupling capacity, Deltap, which measure the probability of forming a new weak bond depending on the number of similar bonds already formed and it is also responsible for the transition. In this paper we show how the model works for a large number of identical molecules and how it can be useful for studying the noise around the centre of the transition where, increasing the degree of cooperativity, i.e. the number n in the well-known Hill equation, the width of the noise increases along with its fractal dimension. A simple relationship between the degree of cooperativity and the parameter Deltap is proposed, suggesting that the cooperativity of real biological transitions is related to the coupling capacity Deltap of the present model.