The use of streptavidin-biotinylglycans as a tool for characterization of oligosaccharide-binding specificity of lectin.

A new rapid and sensitive method for characterizing lectin specificity using streptavidin-biotinylglycans as a tool is presented. This assay is analogous to enzyme immunoassay and takes advantage of the strong, irreversible adsorption of streptavidin to the wells of the chambers of titer plates. A series of streptavidin-biotinylglycans was first coated on a microtiter plate, and then one of six lectins, concanavalin A, wheat germ agglutinin, Phaseolus vulgaris (red kidney bean) erythro-agglutinin, Lens culinaris (lentil) agglutinin, Datura stramoniun agglutinin, or Sambucus nigra (elderberry bark) agglutinin coupled to horseradish peroxidase, was added. After incubation and thorough washing, only the lectin bound to a complementary glycan remained and could be detected and quantified by the peroxidase reaction. It was established that the lectins retained their oligosaccharide-binding specificities after coupling to the peroxidase, that the binding was inhibited by addition of the corresponding sugar inhibitors, and that the color intensity produced by the enzyme reaction is proportional to the amount of lectin-peroxidase bound to biotinylglycan complexed with streptavidin immobilized on the plate. As an example, it was found that the peroxidase-D. stramoniun agglutinin conjugate strongly bound biotinylglycans, GlcNAc3-Man5-R, GalGlcNAc3Man5-R, and GlcNAc3-4Man3-R (R = GlcNAc2-[6-(biotinamido)hexanoyl]-Asn). As little as 10 pmol/ml of lectin was detected. With the growing availability of biotinylglycans, the method should represent a reliable and simple procedure for screening lectin-oligosaccharide recognition qualitatively and quantitatively.     

Chaperonins GroEL and GroES: views from atomic force microscopy.

The Escherichia coli chaperonins, GroEL and GroES, as well as their complexes in the presence of a nonhydrolyzable nucleotide AMP-PNP, have been imaged with the atomic force microscope (AFM). We demonstrate that both GroEL and GroES that have been adsorbed to a mica surface can be resolved directly by the AFM in aqueous solution at room temperature. However, with glutaraldehyde fixation of already adsorbed molecules, the resolution of both GroEL and GroES was further improved, as all seven subunits were well resolved without any image processing. We also found that chemical fixation was necessary for the contact mode AFM to image GroEL/ES complexes, and in the AFM images. GroEL with GroES bound can be clearly distinguished from those without. The GroEL/ES complex was about 5 nm higher than GroEL alone, indicating a 2 nm upward movement of the apical domains of GroEL. Using a slightly larger probe force, unfixed GroEL could be dissected: the upper heptamer was removed to expose the contact surface of the two heptamers. These results clearly demonstrate the usefulness of cross-linking agents for the determination of molecular structures with the AFM. They also pave the way for using the AFM to study the structural basis for the function of GroE system and other molecular chaperones.     

Micropipette suction for measuring piconewton forces of adhesion and tether formation from neutrophil membranes.

A new method for measuring piconewton-scale forces that employs micropipette suction is presented here. Spherical cells or beads are used directly as force transducers, and forces as small as 10-20 pN can be imposed. When the transducer is stationary in the pipette, the force is simply the product of the suction pressure and the cross-sectional area of the pipette minus a small correction for the narrow gap that exists between the transducer and the pipette wall. When the transducer is moving along the pipette, the force on it is corrected by a factor that is proportional to the ratio of its velocity relative to its drag-free velocity. With this technique, the minimum force required to form a membrane tether from neutrophils is determined (45 pN), and the length of the microvilli on the surface of neutrophils is inferred. The strength of this technique is in its simplicity and its ability to measure forces between cells without requiring a separate theory or a calibration against an external standard and without requiring the use of a solid surface.     

拟夏孢锈属（Uredinopsis）新记录种

采到骨状拟夏孢锈（Ｕｒｅｄｉｎｏｐｓｉｓ ｏｓｓｉｆｏｒｍｉｓ Ｋａｍｅｉ）为中国新记录种。     

中药凌霄花的药理学考察

实验结果表明,凌霄花对离体猪冠状动脉条具有抑制收缩的作用;对大鼠血栓形成有抑制作用;能加快红细胞电泳,增加红细胞电泳率,使血液红细胞处于分散状态;对离体孕子宫能显著增强收缩活性,增加收缩频率及增强收缩强度。以上结果可初步说明,凌霄花行血去瘀的作用及其“孕妇慎用”的合理性。凌霄花的３种混淆品（白泡桐花,毛泡桐花,兰考泡桐花）的药理作用与凌霄花不同,故不能作凌霄花使用。     

地衣霉素对细胞膜表面运铁蛋白受体功能的影响

应用抑制糖蛋白Ｎ－糖链合成的地衣霉素处理ＳＭＭＣ－７７２１人肝癌细胞,３Ｈ甘露糖掺入实验显示细胞膜表面糖蛋白Ｎ－糖链的合成受到显著抑制,但细胞膜表面运铁蛋白受体内吞再循环的过程无显明变化,进一步的研究表明受体与运铁蛋白的亲和力亦无改变,但细胞膜表面运铁蛋白受体数减少。结果提示用地衣霉素处理细胞后,在内质网合成的无Ｎ－糖链的运铁蛋白受体影响其运输到细胞膜表面表达。     

红毛五加叶的三萜皂甙

红毛五加（ＡｃａｎｔｈｏｐａｎａｘｇｉｒａｌｄｉｉＨａｒｍｓ）系五加科五加属植物,分布于河北、河南、四川、陕西、甘肃等省,有祛风湿、通关节、强筋骨、治痿痹等功效［１］。其化学成分未见报道。本文报道红毛五加叶的５个三萜皂甙（甙Ⅰ、Ⅱ、Ⅲ、Ⅳ和Ⅴ）的分离鉴定。它们均首次从该属植物发现,其结构如下：　　　　　　　　　　　　　Ｒ１　　　　　　　　　　　　　　Ｒ２　　　　　　　　Ⅰ　ｒｈａｍ－（１→２）－ａｒａ　　　　　Ｈ　　　　　　　Ⅱ　Ｈ　　　ｒｈａｍ－（１→４）－ｇｌｃ－（１→６）－ｇｌｃ　　　　　…     

Big flies, small repeats: the "Thr-Gly" region of the period gene in Diptera

The region of the clock gene period (per) that encodes a repetitive tract of threonine-glycine (Thr-Gly) pairs has been compared between Dipteran species both within and outside the Drosophilidae. All the non- Drosophilidae sequences in this region are short and present a remarkably stable picture compared to the Drosophilidae, in which the region is much larger and extremely variable, both in size and composition. The accelerated evolution in the repetitive region of the Drosophilidae appears to be mainly due to an expansion of two ancestral repeats, one encoding a Thr-Gly dipeptide and the other a pentapeptide rich in serine, glycine, and asparagine or threonine. In some drosophilids the expansion involves a duplication of the pentapeptide sequence, but in Drosophila pseudoobscura both the dipeptide and the pentapeptide repeats are present in larger numbers. In the nondrosophilids, however, the pentapeptide sequence is represented by one copy and the dipeptide by two copies. These observations fulfill some of the predictions of recent theoretical models that have simulated the evolution of repetitive sequences.      

高等植物性别分化研究的某些进展

高等植物性别分化研究的某些进展邵宏波（四平师范学院生物工程研究室吉林四平１３６０００）关键词高等植物,性别分化,基因表达ＳＯＭＥＡＤＶＡＮＣＥＳＩＮＴＨＥＳＥＸＵＡＬＤＩＦＦＥＲＥＮＴＩＡＴＩＯＮＲＥＳＥＡＲＣＨＯＦＨＩＧＨＥＲＰＬＡＮＴＳ￥Ｓｈａｏ...     

番茄种子经激光和咖啡因处理后其当代生物效应的研究

咖啡因具有抑制一些酶的活性,可减弱或消除生物体自身对激光引起的生物效应的修复作用。试验证明咖啡因对番茄种子萌发有强烈的抑制作用,咖啡因和激光复合处理生物效应显著,可降低番茄种子的发芽,并促进番茄果实增大和生长前期速率加快。