Processing of Laminin-5 and Its Functional Consequences: Role of Plasmin and Tissue-type Plasminogen Activator

The laminin-5 component of the extracellular matrices of certain cultured cells such as the rat epithelial cell line 804G and the human breast epithelial cell MCF-10A is capable of nucleating assembly of cell– matrix adhesive devices called hemidesmosomes when other cells are plated upon them. These matrices also impede cell motility. In contrast, cells plated onto the laminin-5–rich matrices of pp126 epithelial cells fail to assemble hemidesmosomes and are motile. To understand these contradictory phenomena, we have compared the forms of heterotrimeric laminin-5 secreted by 804G and MCF-10A cells with those secreted by pp126 cells, using a panel of laminin-5 subunit-specific antibodies. The α3 subunit of laminin-5 secreted by pp126 cells migrates at 190 kD, whereas that secreted by 804G and MCF-10A cells migrates at 160 kD. The pp126 cell 190-kD α3 chain of laminin-5 can be specifically proteolyzed by plasmin to a 160-kD species at enzyme concentrations that do not apparently effect the laminin-5 β and γ chains. After plasmin treatment, pp126 cell laminin-5 not only impedes cell motility but also becomes competent to nucleate assembly of hemidesmosomes. The possibility that plasmin may play an important role in processing laminin-5 subunits is supported by immunofluorescence analyses that demonstrate colocalization of laminin-5 and plasminogen in the extracellular matrix of MCF-10A and pp126 cells. Whereas tissue-type plasminogen activator (tPA), which converts plasminogen to plasmin, codistributes with laminin-5 in MCF-10A matrix, tPA is not present in pp126 extracellular matrix. Treatment of pp126 laminin-5–rich extracellular matrix with exogenous tPA results in proteolysis of the laminin-5 α3 chain from 190 to 160 kD. In addition, plasminogen and tPA bind laminin-5 in vitro. In summary, we provide evidence that laminin-5 is a multifunctional protein that can act under certain circumstances as a motility and at other times as an adhesive factor. In cells in culture, this functional conversion appears dependent upon and is regulated by tPA and plasminogen.     

Genetic analysis of rice CMS-WA fertility restoration based on QTL mapping

 The inheritance of fertility restoration of rice cytoplasmic male sterility of the wild abortive type was studied by means of QTL mapping. The two segregating populations examined showed high frequencies of highly sterile and highly fertile progenies, but a low frequency of partially sterile and partially fertile progenies. The distributions suggested that fertility restoration was mainly controlled by major genes. Based on a linkage map constructed with 57 RFLP and 61 AFLP markers on a B₁F₁ population, composite interval mapping (CIM) revealed that the fertility was restored by the additive effects of two restorer loci located on chromosome 10. One QTL, tightly linked to RFLP marker C1361 in the middle of the long arm of chromosome 10, explained 71.5% of the phenotypic variance. The second QTL was located between RFLP markers R2309 and RG257 on the short arm and explained 27.3% of the phenotypic variance. Similar results were obtained using the simple interval mapping (SIM) methods. Recived: 8 January 1998/Accepted: 22 April 1998     

Shell differentiation and engrailed expression in the Ilyanassa embryo

 We have used in situ hybridization and immunocytochemistry to study the expression of the engrailed-related gene, Ily-en in embryos of the marine mud snail Ilyanassa obsoleta. We find that Ily-en is only expressed in shell gland cells. Only mRNAs localized in the shell gland hybridize to an antisense probe of the Ily-en homeobox. Similarly, only shell gland cells or shell-forming cells are stained by the monoclonal antibody 4D9, which was raised to the engrailed-class protein from Drosophila. Ilyanassa embryos made deficient in vegetal cytoplasm by removing the third polar lobe fail to differentiate an organized external shell. They do however make some randomly oriented internal shell fragments in which Ily-en is expressed. Because Ily-en is expressed in shell gland cells of both normal and lobeless embryos, we conclude that the determinant(s) required for Ily-en expression are not exclusively localized in the polar lobe. Received: 16 October 1997 / Accepted: 9 January 1998     

用膨胀床金属亲和层析从淡菜匀浆液中分离纯化纤维素酶

研究了一种新的膨胀床金属亲和层析技术,即将金属亲和层析结合膨胀床层析,直接从淡菜(Blue mussel)匀浆液中纯化纤维素酶。研究了金属亲和配基种类、pH、离子强度及流速对酶吸附和解吸的影响,确定了酶洗脱条件和介质再生条件。一步可纯化纤维素酶194倍,酶收率达82%。本方法不需要预先去除细胞碎片,而且处理速率比传统层析技术高3～4倍。     

中华鳖生化组成的分析：Ⅱ.背甲,肌肉中矿物元素的组成

本文对中华鳖一龄鳖,二龄鳖和三龄鳖的肌肉与背甲中矿物元素的组成进行了分析,结果表明：中华鳖体内含有２２种以上的矿物元素,具有重要生理功能的硒,锌和铁等微量元素含量丰富,有肌肉中含量分别为：０．５４,３．３２,３６．７;在背甲中含量分别为：１．５０,８．１２,８．８。     

中国株丙型肝炎病毒（HCV）结构区蛋白在昆虫细胞中的表达及加工

利用杆状病毒表达系统在昆虫细胞中表达了完整的中国河北株丙丙型肝炎病毒结构蛋白。免疫印迹实验结果显示,表达产物中有一系列分子量不同、可以与ＨＣＶ抗体阳性病人血清反应的蛋白,表明结构蛋白被宿主细胞蛋白酶切割与加工,相应分别为２０ｋＤ的核心蛋白、３２ｋＤ糖基化的Ｅ１蛋白４０ｋＤ的未糖化的Ｅ２蛋白和７０ｋＤ糖基化的Ｅ２蛋白,另有８０ｋＤ及１００ｋＤ的两组前体蛋白。利用表达产物检测慢性ＨＣＶ感染者血清,发现     

影响水稻纹枯病流行,危害的因子分析

以连作早稻为研究对象,对影响水稻纹枯病发生、危害有关的因子,即品种、施氮肥量、气象因素、为害损失、发病时间、病情程度及药剂等作了系统的定量研究．结果表明,品种间存在抗病性和危害损失程度上的差异;施氮肥量与发病程度关系密切;气象因素中以日均温和雨日频率与病害流行速率关系密切;发病时间与为害损失率相关性不明显,药剂防治效果与控病时间、病情基数有关．     

小麦品种间感染纹枯病的差异及普遍率与严重度的关系

通过人工接菌方法比较了６个小麦主栽品种间感染纹枯病的差异,结果表明,高感类型有阜阳８６１、温麦４号和皖麦１９,中感类型有扬麦１５８、豫麦１８和豫麦２１．各品种苗期病株率反映不出品种感病程度的差异,以灌浆后期的病情指数为标准比较品种间抗感染程度较为适宜．认为寄主生育阶段影响小麦纹枯病的ＩＳ关系．回归分析表明,按内茎和外茎发病程度分级可减少田间调查误差,且省时省工．     

人脊髓前角运动神经元190KD蛋白的初步研究

本研究采用ＳＤＳ凝胶电泳方法从人脊神经前根中分离出人脊髓前角运动神经元特有的蛋白—１９０ＫＤ。将该蛋白作为抗原,免疫ＢＡＬＢ／ｃ小鼠,经杂交瘤技术,获得了抗１９０ＫＤ蛋白的单克隆抗体。免疫细胞化学检测表明,１９０ＫＤ单抗与脊髓灰质前角神经元、前根及肌支发生阳性反应。实验结果提示,１９０ＫＤ蛋白分布在脊髓运动神经元胞体及脊神经的前根和肌支纤维中。