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91.
Haruki Hasegawa Carla ForteIrene Barber Shanon TurnbaughJanelle Stoops Min ShenAi Ching Lim 《Biochimica et Biophysica Acta (BBA)/Molecular Cell Research》2014
Crystalline bodies (CBs) can develop in the endoplasmic reticulum (ER) of antibody-producing cells. Although this phenotype is often reported in association with plasma cell dyscrasias and other hematological disorders, the details of CB biogenesis and CB's roles in pathophysiology remain poorly understood. Using an imaging-based screening method, we identified a secretion-competent human IgG2/λ clone that develops spindle-shaped intracellular crystals in transiently-transfected HEK293 cells upon Brefeldin A treatment. When stably overexpressed from CHO cells, the IgG2/λ clone spontaneously produced spindle-shaped CBs in the ER. Some CBs were released to the extracellular space while remaining enclosed by the membranes of secretory pathway origin. Structural modeling on the variable-region did not uncover prominent surface characteristics such as charge clusters. In contrast, alterations to the constant domain-encoded properties revealed their modulatory roles in CB-inducing propensities and CB morphology. For example, deletion of the entire Fc domain changed the morphology of CBs into thin filaments. Elimination of an N-linked glycan by a N297A mutation promoted Russell body biogenesis accompanied by marked reduction in IgG secretion. Isotype class switching from the original IgG2 to IgG1 and IgG4 changed the crystal morphology from spindle-shaped to long needle and acicular shaped, respectively. The IgG3 version, in contrast, suppressed the CB formation. Either the HC or LC alone or the Fc-domain alone did not trigger CB biogenesis. An IgG's in vivo crystal morphology and crystallization propensity can thus be modulated by the properties genetically and biochemically encoded in the HC constant region. 相似文献
92.
喉癌差异表达cDNA序列的分离与初步鉴定 总被引:12,自引:0,他引:12
分离和克隆人喉癌中新的相关基因将有助于提示喉癌的易感性与癌变机制。运用mRNA差异显示法对2例成人喉癌组织及配对癌旁正常组织的基因表达进行研究,分离到35个差异显示片段;用反向Northern点杂交筛选到6个差异片段,经克隆、测序和匹配分析,得12条不同cDNA序列,其中4条为新基因序列,另外8条与已知基因高度同源。将12条cDNA序列固定在膜上,用来自喉癌和配对正常组织的总cDNA探针与其杂交和 相似文献
93.
The aquatic monocot Hydrilla verticillata (L.f.) Royle is a well-documented facultative C4 NADP-malic enzyme species in which the C4 and Calvin cycles operate in the same cell with the specific carboxylases confined to the cytosol and chloroplast, respectively. Several key components had already been characterized at the molecular level, thus the purpose of this study was to begin to identify other, less obvious, elements that may be necessary for a functional single-cell C4 system. Using differential display, mRNA populations from C3 and C4 H. verticillata leaves were screened and expression profiles compared. From this study, 65 clones were isolated and subjected to a customized macroarray analysis; 25 clones were found to be upregulated in C4 leaves. Northern and semi-quantitative RT-PCR analyses were used for confirmation. From these screenings, 13 C4 upregulated genes were identified. Among these one encoded a previously recognized C4 phosphoenolpyruvate carboxylase, and two encoded distinct pyruvate orthophosphate dikinase isoforms, new findings for H. verticillata. Genes that encode a transporter, an aminotransferase and two chaperonins were also upregulated. Twelve false positives, mostly housekeeping genes, were determined from the Northern/semi-quantitative RT-PCR analyses. Sequence data obtained in this study are listed in the dbEST database (DV216698 to DV216767). As a single-cell C4 system that lacks Kranz anatomy, a better understanding of how H. verticillata operates may facilitate the design of a transgenic C4 system in a C3 crop species.Srinath K. Rao and Hiroshi Fukayama contributed equally to this study. 相似文献
94.
95.
A proteolytic susceptibility index was proposed to evaluate the reaction performance of different food by-products with subtilisin. Whey, salmon muscle and feather keratine were hydrolyzed at the same peptide bond concentration at 50 °C and pH 8.0 with different subtilisin concentrations. The logarithmic equation P = 1/b ln(abt+1) was fitted to estimate the kinetic constants a and b. The a/b ratio was proposed as a proteolytic susceptibility index, which was correlated with the chemical structure and conformation of protein sources in increasing order of feather keratin, salmon muscle and whey proteins with corresponding a/b values of 0.1, 8.2 and 14.4 mM2/min, respectively. The methodology proposed in this work can be used to evaluate the proteolytic susceptibility of different protein by-products to different proteases and to evaluate the changes in proteolytic susceptibility after treatment. 相似文献
96.
Pairs of neurons were differentially stained by intracellular injection of the divalent cations, nickel and cobalt. Pairs of neuronal tracts could be similarly labelled following immersion of their axon bundles in the markers described. Use of nickel and a 4 : 6 mixture of cobalt and nickel according to the protocols presented here revealed the morphology of cricket neuron processes as thin as 1 μm in diameter and yielded a favourable contrast between the cells marked, as well as with respect to the background of unstained tissue. 相似文献
97.
随着基因芯片的技术的推广,越来越多的表达数据需要被处理和分析.利用这些表达数据提取基因调控矩阵从而构建基因网络是一个重要的问题.通过线性微分方程模型可以初步构建基因网络,了解网络结构,提取最显著的信息.然而由于分子生物学的条件限制或者数据来源的限制,导致实验数据不充分,使方程组无解.本文使用三次样条方法,对26例临床、病理资料完备的具有淋巴结转移的乳腺癌基因表达数据进行插值处理,使表达数据满秩,从而使用最小二乘法解出加权矩阵,构建初步的表达基因调控网络.通过对构建的基因网络的初步分析表明:乳腺癌转移的形成是由多基因异常引起多条传导通路异常,致使细胞恶性转化的结果,这与生物学上公认的看法是相一致的.因此,利用此线性模型方法对基因表达谱进行分析兵有一定可行性,在认识乳腺癌转移机制,乳腺癌诊断和治疗方面具有一定的理论和应用价值. 相似文献
98.
Effects of sorbitol addition on the action of free and immobilized hydrolytic enzymes in organic media 总被引:4,自引:0,他引:4
Triantafyllou AO Wehtje E Adlercreutz P Mattiasson B 《Biotechnology and bioengineering》1995,45(5):406-414
The effect of the addition of sorbitol on the activity and stability of enzymes was examined by monitoring transesterification reactions performed in organic media at various water activities (a(w) = 0.08 to 0.97). Lipases from Chromobacterium viscosum and Candida rugosa immobilized on celite, and chymotrypsin, free or immobilized on celite, were used. When the sorbitol-containing enzymes were employed, higher reaction rates and less hydrolysis were observed. Immobilization of chymotrypsin resulted in high activity and operational stability, while the nonimmobilized enzyme was stable only in the presence of sorbitol. The activity of all preparations diminished after washing them with pyridine to remove sorbitol. Furthermore, severe stability problems occurred in the preparations lacking sorbitol. Sorbitol treatment, even after removal of the sorbitol itself, improved the activity of nonimmobilized chymotrypsin relative to the washed control. On the other hand, washing to remove sorbitol had a negative effect on the activity of both coimmobilized lipase and coimmobilized chymotrypsin. Addition of a substrate analogue, N-acetyl-L-phenylalanine, to chymotrypsin yielded a preparation that exhibited higher activity than both the control and its sorbitol-containing counterpart. Differential scanning calorimetry measurements revealed that the chymotrypsin-sorbitol complex was stable against thermal denaturation, undergoing transition at a high temperature (89 degrees C). The transition temperatures of the substrate-containing chymotrypsin and of the control were identical (72 degrees C). (c) 1995 John Wiley & Sons, Inc. 相似文献
99.
Significant progress has been made in elucidating the mechanisms used by plants to recognize pathogens and activate “immune” responses. A “first line” of defense can be triggered through recognition of conserved Pathogen or Microbe Associated Molecular Patterns (PAMPs or MAMPs), resulting in activation of basal (or non-host) plant defenses, referred to as PAMP-triggered immunity (PTI). Disease resistance responses can also subsequently be triggered via gene-for-gene type interactions between pathogen avirulence effector genes and plant disease resistance genes (Avr-R), giving rise to effector triggered immunity (ETI). The majority of the conceptual advances in understanding these systems have been made using model systems, such as Arabidopsis, tobacco, or tomato in combination with biotrophic pathogens that colonize living plant tissues. In contrast, how these disease resistance mechanisms interact with non-biotrophic (hemibiotrophic or necrotrophic) fungal pathogens that thrive on dying host tissue during successful infection, is less clear. Several lines of recent evidence have begun to suggest that these organisms may actually exploit components of plant immunity in order to infect, successfully colonize and reproduce within host tissues. One underlying mechanism for this strategy has been proposed, which has been referred to as effector triggered susceptibility (ETS). This review aims to highlight the complexity of interactions between plant recognition and defense activation towards non-biotrophic pathogens, with particular emphasis on three important fungal diseases of wheat (Triticum aestivum) leaves. 相似文献
100.