Nonenzymatic chromophore attachment in biliproteins: conformational control by the detergent Triton X-100

While chromophore attachment to alpha-subunits of cyanobacterial biliproteins has been studied in some detail, little is known about this process in beta-subunits. The ones of phycoerythrocyanin and C-phycocyanin each carry two phycocyanobilin (PCB) chromophores covalently attached to cysteins beta84 and beta155. The differential nonenzymatic reconstitution of PCB to the apoproteins, PecA, PecB, CpcA and CpcB, as well as to mutant proteins of the beta-subunits lacking either one of the two binding cysteins, was studied using overexpression of the respective genes. PCB adds selectively to Cys-84 of CpcA, CpcB, PecA, and PecB, but the bound chromophore has a nonnative configuration, and in the case of CpcA, is partly oxidized to mesobiliverdin (MBV). The oxidation is independent of thiols but can be suppressed by ascorbate. The addition to Cys-beta84 is suppressed in the presence of detergents like Triton X-100, in favor of an addition to Cys-beta155 yielding the correctly bound chromophore. Triton X-100 also inhibits oxidation of the chromophore during addition to CpcA. The effect of Triton X-100 was studied on the isolated components of the reconstitution system. Absorption, fluorescence and circular dichroism spectra indicate a major conformational change of the chromophore upon addition of the detergent, which probably controls the site selectivity of the addition reaction, and inhibits the oxidation of PCB to MBV.     

Brain-specific potential guanine nucleotide exchange factor for Arf, synArfGEF (Po), is localized to postsynaptic density

We cloned from a rat brain cDNA library a novel cDNA and named it a potential synaptic guanine nucleotide exchange factor (GEF) for Arf (synArfGEF (Po)) (GenBank Accession no. AB057643) based on its domain structure and localization. The cloned gene was 7410 bases long with a 3585-bp coding sequence encoding a protein of 1194 amino acids. The deduced protein contained a coiled-coil structure in the N-terminal portion followed by Sec7 and Plekstrin homology (PH) domains. Thus, the protein was a member of the Sec7 family of proteins, GEFs. Conservation of the ADP-ribosylation factor (Arf)-binding sequence suggested that the protein was a GEF for Arf. The gene was expressed specifically in the brain, where it exhibited region-specific expression. The protein was highly enriched in the postsynaptic density (PSD) fraction prepared from the rat forebrain. Uniquely, the protein interacted with PSD-95, SAP97 and Homer/Vesl 1/PSD-Zip45 via its C-terminal PDZ-binding motif and co-localized with these proteins in cultured cortical neurons. These results supported its localization in the PSD. The postsynaptic localization was also supported by immunohistochemical examination of the rat brain. The mRNA for the synArfGEF was also localized to dendrites, as well as somas, of neuronal cells. Thus, both the mRNA and the protein were localized in the postsynaptic compartments. These results suggest a postsynaptic role of synArfGEF in the brain.     

鱼腥藻PCC7120藻蓝蛋白α亚基体内重组研究

为了研究藻蓝蛋白α亚基的生物合成途径,通过构建相容的3种重组质粒pETDuet-cpcA、pCOLADuet-cpcE-cpcF和pACYCDuet-ho1-pcyA,将裂合酶基因cpcE和cpcF、血红素氧化酶基因ho1、藻蓝胆素合成酶基因pcyA和脱辅基藻蓝蛋白α亚基基因cpcA共同转入大肠杆菌BL21(DE3)。通过色素蛋白锌电泳和光谱检测表明产生了生物活性的CpcA-PCB。成功实现了大肠杆菌内藻蓝蛋白α亚基84位半胱氨酸残基与PCB的连接。而在裂合酶基因cpcE和cpcF不转入大肠杆菌的情况下,大肠杆菌内只有0.2%的CpcA-PCB产生。以上研究为进一步在大肠杆菌内合成天然的藻蓝蛋白奠定了基础。     

西花蓟马取食不同豆科蔬菜的实验种群生命表

西花蓟马Frankliniella occidentalis(Pergande)是传入我国的重要入侵害虫,在贵阳地区对豆科蔬菜为害十分严重。在25℃条件下,组建了西花蓟马取食大豆叶片、豇豆叶片、四季豆叶片和四季豆豆荚时的实验种群生命表。结果表明:西花蓟马未成熟期取食四季豆豆荚时存活率最高,取食其它3种豆科蔬菜时存活率相差不大。西花蓟马取食四季豆豆荚时内禀增长力最高为0.1626,取食豇豆叶片时最低为0.0834,净增值率也是取食四季豆豆荚时最高,取食大豆叶片时最低,表明四季豆豆荚最有利于西花蓟马的生长发育和繁殖。在不同豆科蔬菜上的稳定年龄组配中未成熟期所占的比例很大,而成虫期所占的比例相对较小。     

东方田鼠胚胎成纤维细胞的分离培养及生物学特性

目的对影响东方田鼠胚胎成纤维细胞（MfEF）分离和培养的因素进行探索,并观察其生物学特性。方法取室内繁殖饲养不同胎龄的东方田鼠胚胎分离成纤维细胞,通过原代和继代培养,分析比较不同胎龄、不同血清浓度、不同胰蛋白酶浓度等因素对MfEF分离及培养的影响,观察MfEF的生长形态及其生物学特性。结果 MfEF为贴壁型生长,细胞形态多样,呈梭形、不规则多边形;采用0.125%的胰蛋白酶室温消化12～13 d胚胎组织5 min,以DMEM培养基添加15%小牛血清分离培养MfEF的效果最佳;MfEF2～7代增殖最旺盛。结论获得了实验室分离、培养MfEF的有效方法 ,为进一步深入研究东方田鼠抗日本血吸虫机制以及开展不同动物成纤维细胞间比较研究奠定了基础。     

丙烯对柿果实采后细胞壁物质代谢和几种生理指标的影响

丙烯处理的柿果实,其软化进程显著加快,呼吸速率和乙烯释放高峰提前,且峰值升高,细胞壁各组分的代谢速度加快,柿果实中多聚半乳糖醛酸酶（PG）和纤维素酶（Cx）活性提高,且高峰提前。     

Erk is involved in the differentiation induced by diallyl disulfide in the human gastric cancer cell line MGC803

Diallyl disulfide (DADS) is a major constituent of garlic. Previously, we found that DADS both inhibited proliferation in human gastric cancer cells in vitro and in vivo, and induced G2/M arrest. In this study, we investigated whether this differentiation effect was induced by DADS in human gastric cancer MGC803 cells, and whether it was related to an alteration in ERK activity. The results showed that the growth of MGC803 cells was inhibited by DADS. Cells treated with DADS displayed a lower nucleocytoplasmic ratio and tended to form gland and intercellular conjunction structures. The ConA-mediated cell agglutination ratio and cells’ ALP specific activity decreased. In MGC803 cells, dye transfer was limited to a few cells neighbouring the dye-injected cell and to a depth of 1–2 layers beneath the scrape site. However, after treatment with DADS, the LY (Lucifer Yellow) was transferred to several cells immediately neighbouring the microinjected cell and to a depth of 2–4 cell layers from the scrape site. This indicated that DADS induced differentiation in MGC803 cells. Western blot analysis revealed that although DADS did not influence the quantity of ERK1/2 protein expressed, it did decrease its phosphorylation in a concentration-dependent manner, compared with the controls. At 30 mg·L⁻¹, DADS inhibited the activation of ERK1/2 in 15–30 min. These results suggested that the DADS-induced differentiation of MGC803 cells involved an alteration of the ERK1/2 signaling pathway.     

A NEW GENUS OF EUGALEASPIDIFORMS (AGNATHA: GALEASPIDA)FROM THE LUDLOW, SILURIAN OF QUJING, YUNNAN,SOUTHWESTERN CHINA

记述了云南曲靖潇湘区关底组(志留系罗德洛统)真盔甲鱼类一新属、新种——长孔盾鱼(Dunyu longiforus gen.et sp.nov.),丰富了以梦幻鬼鱼为代表的潇湘脊椎动物群的组成.该新属具真盔甲鱼科的鉴别特征:后眶上管和中背管二者连续过渡,后端两侧辏合呈“U”字形,侧背管仅发出3条侧横管.新属内角缺如,这一特征过去在真盔甲鱼科中仅见于真盔甲鱼属.新属区别于真盔甲鱼属的主要特征有:角向后方延伸,呈棘状或叶状;头甲最宽处位于角末端之前,宽长比小于1.1;中背孔末端超越眶孔后缘连线.基于对秀山真盔甲鱼头甲标本的重新观察,将秀山真盔甲鱼归入盾鱼属.新种区别于秀山盾鱼(Dunyu xiushanensis)的特征有:个体较大,头甲长大于宽;角呈棘状;眶孔位置相对靠前,头甲眶前区与眶后区的长度之比小于0.75;多边形崮状瘤点,较大,长可超过2 mm.     

一株抗万古霉素耐药菌的抗生素产生菌AR1148的鉴定

从土壤中分离得到一株放线菌AR1148,其代谢产物对万古霉素耐药肠球菌有较明显的抑菌活性。该菌株的基内菌丝无横隔,气生菌丝丰茂,分枝较好,孢子椭圆形,表面光滑。菌丝细胞壁工型,含有L-2,6-二氨基庚二酸（LL-DAP）和甘氨酸。菌株AR1148归属于链霉菌属金色类群。根据16SrDNA序列分析,该菌株与现有种差异明显,聚类在不同的分支。定名为Streptomyces sp．AR1148。