Fluorescence studies of chicken liver fatty acid synthase. Segmental flexibility and distance measurements

The 4'-phosphopantetheine of chicken liver fatty acid synthase was specifically labeled with the fluorescent substrate analog coenzyme A 6-[7-nitrobenz-2-oxa-1,3-diazol-4-yl]aminohexanoate at low salt concentrations. A serine at the active site of the thioesterase was specifically labeled with the fluorescent compounds 6-[7-nitrobenz-2-oxa-1,3-diazol-4-yl]aminopentylmethylphosphono fluoridate and/or pyrenebutyl methylphosphonofluoridate. Dynamic anisotropy measurements indicate the thioesterase has considerable segmental flexibility, whereas the fluorescent labeled 4'-phosphopantetheine does not display detectable local or segmental flexibility. Fluorescence resonance energy transfer measurements indicate that the distance between the fluorescent label at the end of the 4'-phosphopantetheine and NADPH bound to the beta-ketoacyl reductase or enoyl reductase site on the same polypeptide chain is essentially the same, approximately 38 A. The two types of reductases were distinguished by specifically blocking enoyl reductase with pyridoxal 5'-phosphate. No significant energy transfer occurs between sites on different polypeptide chains so that the distances must be greater than 55 A. The distance between the serine on the thioesterase and the 4'-phosphopantetheine on the same polypeptide is 48 A; again no interpolypeptide chain energy transfer was observed. The distance between the serines of the two thioesterases within a fatty acid synthase molecule is greater than 56 A. The monomeric enzyme obtained at 1 degree C does not have beta-ketoacyl synthase and reductase activities. Also fluorescent titrations indicate NADPH is not bound to beta-ketoacyl reductase in monomeric enzyme. The addition of potassium phosphate to the monomers at 1 degree C rapidly dimerizes the enzyme and restores the beta-ketoacyl reductase activity. The beta-ketoacyl synthase activity is slowly restored when the dimer is raised to room temperature. The results obtained suggest that relatively large conformational changes may be part of the catalytic cycle.     

神农香菊干花净油成分的研究

神农香菊全草提取的香浸膏香气浓郁独特,可用于调配多种高档化妆用香精,也可直接用于饮料中,在香精香料工业中具有较大的实用价值。本文首次报告了我们用毛细管气相色谱仪和色谱/质谱/计算机联用分析仪,对神农香菊干花净油成分进行分析的初步结果。鉴定出的已知成分包括脂肪族类,含氧或非含氧的单萜及倍半萜类化合物32个。     

蝮蛇毒抗凝血活酶组份及几种蛇毒粗毒对人红细胞和血小板的作用

蝮蛇毒抗凝血活酶组分及蝗蛇毒、圆斑蝰蛇毒和眼镜蛇毒粗毒,不仅能够水解血浆中的磷脂,而且还能水解完整人红细胞膜和完整人血小板膜上的磷脂。但是五步蛇毒和金环蛇毒粗毒却不能水解完整人血小板膜上的磷脂。 扫描电镜观察表明,由于抗凝血活酶组份和几种蛇毒粗毒的作用,人红细胞和人血小板的形态发生了巨大的变化;人红细胞由正常的双圆盘形变成带刺的小球,人血小板的外形变成蜂窝状。     

叶潜蛾科一个新种——黄皮叶潜蛾(鳞翅目:叶潜蛾科)

叶潜蛾属Phyllocnistis Zeller已从细蛾科Gracillariidae分立出来,另独立成为叶潜蛾科Phyllocnistidae(Kloct and Hincks,1972)。根据Imms(1977)记载,该科在全世界约有50多种。我们最近查阅文献,该科已有80多种。但本科在我国记载的种类仅有柑桔叶潜蛾Phyllocnistis cilrella Stainton及杨银叶潜蛾Phyllocnistis saligna Zeller。它们的食     

低温对不同耐寒力的黄瓜幼苗子叶的各细胞器中NAD~+-苹果酸脱氢酶的影响

NAD~+-MDH在黄瓜子叶中的定位是细胞溶质中占总活性的55～59％,线粒体为38～35％,叶绿体为7％。其同工酶谱亦为细胞溶质中带数最多,全青为5条,粤早3号为4条,线粒体和叶绿体均为1条,品种间无明显差异。黄瓜幼苗随低温胁迫的加剧,伤害逐步加重,子叶电解质渗出率明显增加,NAD~+-MDH活性亦不断下降,其中叶绿体的NAD~+-MDH对低温最敏感,1±1℃处理就能反映品种间耐寒力的差异。叶绿体和线粒体的NAD~+-MDH同工酶对低温的反应与活性变化一致,谱带数没有差异,只是活性降低。细胞溶质部分酶带较多,各条酶带对低温的反应不同。     

用PFU法研究微型生物群集过程中数据的处理

根据MacArthur-Wilson的岛屿区系平衡模型S_t=S_(eq)(1-e~(GT)),可以从野外生态效应试验和室内毒性试验中,提出3个功能参数(S_(eq)、G、t_(90％))进行比较。本文提出两种计算方法:复合梯形法和最小二乘法,后者已在计算机上实现了BASIC计算程序。从数学理论上论证,最小二乘法误差较小,但如果实验布局合理,两种计算方法能得到十分一致的结果。实验模型是否符合理论模型,可以用统计学上的拟合差异度检验法来检验。     

抗人肺癌细胞单克隆抗体的制备及其特性的研究

用人肺鳞癌细胞LTEP-78细胞系免疫Blab/c小鼠获得3株抗人肺癌细胞的单克隆抗体杂交瘤系。其中BLTI-01株经六次克隆化培养,体外传代8个月以上。BLTI-01与白细胞抗原及血型抗原基本上无交叉反应;与骨髓细胞无交叉反应;与癌胚抗原和胎甲球蛋白不相关;与肺鳞癌、肺腺癌细胞系及部分其它肿瘤细胞呈阳性反应。     

苯胺嘧啶衍生物X-9通过下调整合素β1表达抑制肝细胞癌迁移侵袭

整合素在许多肿瘤细胞中高表达,并且参与肿瘤细胞的侵袭转移。在肝细胞癌中,整合素β1被报导高表达,并促进肿瘤细胞的侵袭。目前,对于整合素的表达调控癌细胞机制以及干预其表达进而抑制肿瘤细胞转移的研究较少。本研究探讨利用小分子化合物抑制整合素表达来抑制肿瘤细胞迁移和侵袭的可能。首先,对临床肝癌细胞患者癌组织和癌旁组织中的整合素β1的表达进行检测,发现其在癌组织中的表达显著高于癌旁组织（P<0.05）。对TCGA肿瘤数据库的生物信息学分析结果同样显示,整合素β1的高表达与肝癌的分期（P=0.019）和预后（P=0.013）相关。通过筛选发现,苯胺嘧啶衍生物X09可以抑制肝癌细胞中整合素β1的mRNA和蛋白质的表达（P<0.01）。细胞划痕愈合实验和细胞穿孔实验结果显示,苯胺嘧啶衍生物X-9能够抑制肝癌细胞的迁移和侵袭（P<0.01）。进一步的研究证实,在肝癌细胞中外源表达整合素β1可以逆转X-9对肝癌细胞迁移和侵袭的抑制;而在敲低整合素β1的细胞中,X-9对细胞的迁移和侵袭的抑制被消除。因此,鉴定出苯胺嘧啶衍生物X-9可以通过下调整合素β1表达,进而抑制肝癌细胞的迁移和侵袭。     

Characterization of newly isolated monoclonal antibodies against MHC of a Japanese wild mouse

We have already developed nine B10.MOL congenic strains carrying H-2 haplotypes derived from Japanese wild mice, Mus musculus molossinus, with the C57BL/10 genetic background. To obtain monoclonal antibodies against the H-2 antigen of the Japanese wild mouse, we carried out cell fusion using spleen cells from the animal immunized with one of the B10.MOL strains, B10.MOL-SGR (H-2 ^wm7). As a result, 19 hybridomas producing monoclonal antibodies were produced. Analysis with the intro-H-2 recombinants derived from B10.MOL-SGR indicated that 8 of them reacted with the class I and II with the class II molecule. The class I antibodies were tested for their cross -reactivities on wild mice and on the panels of standard inbred and B10.MOL strains. Most of the antibodies reacted with both the Japanese wild mice and the other subspecies, including standard inbred, while two antibodies highly specific for the donor H-2K region reacted with only three wild-derived mice, two M. m. molossinus from Anj o and Shizuoka, Japan, and one M. m. domesticus from Pigeon, Canada. In addition, all of the other four antibodies reactive with the K antigen of B10.MOL-SGR also reacted with the same three wild mice. The wild mice belonging to different subspecies might share very similar H-2K antigenic determinants in spite of their genetic and geographical remoteness.     

聚乙二醇处理大豆种子子叶中几种酶活性和可溶性蛋白含量的变化

聚乙二醇处理的大豆种子的异柠檬酸裂解酶、苹果酸脱氢酶、过氧化氢酶、超氧物歧化酶、酸性磷酸酶、碱性磷酸酶的活性明显高于受低温吸胀冷害的种子子叶的活性,相关的酶活性协同地增长,而蛋白质的含量没有明显的变化。这些酶活性的提高可能是渗透调控处理对细胞膜系统修补的结果。