Identification of Three Novel Unique Proteins in Seed Oil Bodies of Sesame

Graduate Institute of Agricultural Biotechnology, National Chung-HsingUniversity, Taichung, Taiwan 40227, ROC Plant seeds store triacylglycerolsin discrete organelles called oil bodies. An oil body preservesa matrix of triacylglycerols surrounded by a monolayer of phospholipidsembedded with abundant structural proteins termed oleosins andprobably some uninvestigated minor proteins of higher molecularmass. Three polypeptides of 27, 37, and 39 kDa (temporarilydenominated as Sopl, Sop2, and Sop3) were regularly co-purifiedwith seed oil bodies of sesame. Comparison of amino acid compositionindicated that they were substantially less hydrophobic thanthe known oleosins, and thus should not be aggregated multimersof oleosins. The results of immuno-recognition to sesame proteinsextracted from subcellular fractions of mature seeds, varioustissues, and oil bodies purified from different stages of seedformation revealed that these three polypeptides were uniqueproteins gathered in oil bodies, accompanying oleosins and triacylglycerols,during the active assembly of the organelles in maturing seeds.Both in vivo and in intro, immunofluorescence labeling usingsecondary antibodies conjugated with FITC (fluorescein isothiocyanate)confirmed the localization of these three polypeptides in oilbodies. ¹To whom correspondence should be addressed     

Oxidative Damage to Proteins, Lipids, and DNA in Cortical Brain Regions from Patients with Dementia with Lewy Bodies

Abstract: Dementia with Lewy bodies (DLB) forms the second most common pathological subgroup of dementia after Alzheimer's disease. The present study compares the levels of oxidative damage to proteins, lipids, and DNA bases in cortical brain areas from patients with DLB with levels in matched control tissues. Overall, there was a trend for protein carbonyl levels to be increased in all areas, but a significant difference was found only in the parietal and temporal lobes. No differences were observed in the levels of lipid peroxidation. Measurement of products of damage to DNA bases showed increased levels of thymine glycol, 8-hydroxyguanine, 2,6-diamino-4-hydroxy-5-formamidopyrimidine, 5-hydroxycytosine, 5-hydroxyuracil, 5-hydroxymethyluracil, and xanthine. Xanthine levels were increased in the DLB group in the parietal, occipital, and temporal lobes, indicating that peroxynitrite or other deaminating species may be involved. The finding of increased protein carbonyls and increased DNA base products in cortical regions from DLB patients indicates that oxidative stress may play a role in DLB.     

新疆裂叶苔科11种植物孢子、弹丝和油体的形态观察

为了完善有关裂叶苔科植物孢子、弹丝和油体的研究,为该科植物的分类提供详细的资料,利用光学显微镜和扫描电镜观察裂叶苔科（Lophoziaceae）11种植物的孢子、弹丝和油体的形态特征,其中有6种具备了孢子和弹丝的形态特征,9种具备了油体的形态特征。结果表明：(1)裂叶苔科的6种植物的孢子为球形、近球形和椭圆形;颜色为褐色、红褐色和黄褐色;直径在10～16μm,属于小型孢子（MI）;近极面向内凹,纹饰为疣状、棍棒状、刺状、颗粒状和短棒状。(2)弹丝均为紧密双螺旋状,两端圆钝,表面平滑或具稀的颗粒2种。(3)裂叶苔科的9种植物的油体形状为球形、椭圆形、球形或卵形,直径在3～10μm。本研究对这11种植物的孢子、弹丝和油体进行了详细的描述并附图示,为裂叶苔科植物的系统分类、演化研究提供基础资料。     

Seed Storage Proteins in Solanaceae and Cucurbitaceae Species

Electrophoretic analyses of non-reduced and reduced seed storage proteins from Solanaceae and Cucurbitaceae species and cultivars were performed. High molecular disulfide bonded complexes between intermediary subunits of 11S globulins previously detected in Capsicum annuum cultivars, were found in Solanum melongena cultivars as well. The data obtained might be used for further elucidation of peculiarities of the 11S globulins in dicotyledonous plants.     

Ultrastructural Characterization of Oil Bodies in Different Coffea Species

In the coffee seed, the lipid component known as coffee oil is stored in the endosperm tissue as an energy reserve for germination and post-germination growth. This coffee constituent is present in the form of subcellular spherical oil bodies (“oleosomes”) in a typical size range of 0.2–2.5 μm. These particles are filled with an osmiophilic matrix of triglycerides, delimited by a single protein membrane, typical of oleaginous plant tissues. The object of this study is to characterize the morphology and distribution of oil bodies in different coffee species. In particular, we studied Indian samples of Coffea arabica, C. canephora, C. liberica, C. stenophylla and C. salvatrix. After appropriate fixation and preparation, the samples were examined and oil bodies characterized by optical microscopy and transmission electron microscopy. Oil bodies morphology, tissue distribution and size distribution were determined and several features of these subcellular structures were observed and discussed for the first time in the framework of a coffee inter-species comparative study.     

种子的贮油细胞器——油体及其蛋白

生物柴油作为一种可再生的替代能源,因其良好的环境效应而倍受关注.植物油是生产生物柴油的主要原料.植物油作为最经济、最有利于种子传播和萌发生长的能量和碳源物质,贮存和积累于植物种子的贮油细胞器-油体之中.本文综述了植物种子油体的亚细胞结构、化学组成及其功能、生物合成与降解等的研究进展.     

种子的贮油细胞器——油体及其蛋白

生物柴油作为一种可再生的替代能源, 因其良好的环境效应而倍受关注。植物油是生产生物柴油的主要原料。植物油作为最经济、最有利于种子传播和萌发生长的能量和碳源物质, 贮存和积累于植物种子的贮油细胞器——油体之中。本文综述了植物种子油体的亚细胞结构、化学组成及其功能、生物合成与降解等的研究进展。     

Different Types of Major Storage Seed Proteins in Fagaceae Species

The seed proteins of Fagus sylvatica, Quercus Hex and Q. robur,of the Fagaceae family, were extracted by the sequential methodof Koie and Nielsen (1977). The storage proteins of Fagns werefound to be globulins as has previously been shown in Castanea,a member of the same family. Quercus was found to store glutdins.The amino acid composition and the preliminary characterizationby electro-phoresis of Fagus globulins and Quercus glutelins,suggest that both contain proteins similar to the 1 IS globulinsof leguminous plants. Both genera showed considerable differencesin their albumins. Prolamins were found only in Fagus. Key words: Fagaceae, seeds, storage protein     

杉木雌配子体与后胚发育过程中核酸、蛋白质和类脂的消长

在杉木胚胎分化期至成熟期,每个雌配子体总核酸和DNA,RNA含量在初期增加,后期则随着胚的分化发育逐渐下降,而蛋白质和类脂则一直上升。每胚总核酸、DNA,RNA则相应增加,而蛋白质、类脂的含量和干重亦逐渐增加。在胚分化早期RNA的迅速合成与细胞的分化及器官形成有关。但以胚干重为单位的DNA、RNA含量却随着胚的发育而有所减少;蛋白质含量先增加,至成熟后才下降。授粉前的胚珠,以及雌配子体、胚中都发现有凝集素存在。     

Seed Storage Behavior and Seed Germination of Nine Species of Lauraceae from Yunnan,China

Seed banking following internationally agreed standards is an important way for preserving collections of wild plant species ex situ; but this method is not suitable for desiccation sensitive species. Lauraceae comprehends some of the dominant species in the evergreen broadleaved forest in the south of China and contains many species both of ecological and economical importance. However, study on seed biology such as germination and desiccation tolerance of this family is scarce. Seeds of 9 species from 5 genera of this family were collected and their dormancy status and germination requirement were studied; also their desiccation tolerance were determined using a modified 100 seed test. The results showed that seeds of Cinnamomum camphora probably have intermediate physiological dormancy; seeds of Actinodaphne forrestii, Actinodaphne obovata, Cinnamomum migao, Lindera metcalfiana var. dictyophylla, Lindera communis and Neolitsea polycarpa are non deep physiological dormant; Seeds of Cinnamomum burmannii and Phoebe glaucophylla may have no or negligible dormancy. All 9 species lost seed viability after desiccated to 286%-716% moisture content while still retained considerable viability with moisture content ranged from 1732% to 4487% after moist storage; thus seeds of the 9 species are all desiccation sensitive and can not be stored at the conventional seed bank conditions.     

Comparative Electrophoretic Studies of the Seed Proteins of Certain Species of Brassica and Sinapis

The saline-soluble seed proteins of Brassica campestris var.autumnalis, B. nigra, B. oleracea, B. campestris var. rapa andSinapis alba have been investigated using serological methodsand detection of enzymes after electrophoresis in starch gels.These methods were used to determine the generic status of Sinapisalba and the varietal or specific status of B. campestris var.rapa. The results of protein analysis agree with establishedtaxonomy and suggest that both methods help to resolve sometaxonomic problems.     

Protein Bodies from Buckwheat Seed Cotyledons: Isolation and Characteristics

Protein bodies were isolated from cotyledons of dry buckwheatseeds by homogenization in acetone with subsequent purificationin a 1.26 g cm^–3 to 1.53 g cm^–3 linear density gradientof a mixture of acetone with CCI₄. The purified fraction ofprotein bodies with globoids (PB I) had a buoyant density of1.48–1.51 g cm^–3 and was intact according to microscopicdata. Localization of hydrolytic enzymes and proteinase inhibitorsin the PB I fraction and in the fraction of the cytoplasm andmembrane material (CMM) was studied. It was shown that acidhydrolytic enzymes, such as aspartic proteinase, carboxypeptidase,acid phosphatase, -D-mannosidase and N-acetyl-ß-glucosaminidase,as well as chymotrypsin and trypsin inhibitors were predominantlylocalized in the PB I. BAPAase and SH-activated caseinase activitieswere equally distributed between the PB I and CMM fraction.The activities of leucine aminopeptidase and SH-independentcaseinase were noticeably predominant in the CMM fraction. Key words: Buckwheat, subcellular fractionation, protein bodies, hydrolases     

Comparative Electrophoretic Studies of the Seed Proteins of Certain Amphidiploid Species of Brassica

The saline-soluble seed protein extracts of two varieties ofBrassica nigra, B. Oleracea, B. campestris, B. carinata, B.juncea, and B. napus were investigated. The ß-galactosidases,ß-glucosidases, and esterases were detected afterelectrophoresis of the protein extracts in strach gels. Serologicalmethods were also applied to some of these extracts. The methodswere used in order to investigate the possibility of amphidiploidywithin the genus. The analysis of the seed proteins of suggestedhybrids with references to those of their putative parents isreported in this paper and gives evidence to support the suggestionof hybridization.     

Lipins, Lipids and Nuclear Envelope Structure

The lipid composition of biological membranes is crucial for many aspects of organelle function, including growth, signalling, and transport. Lipins represent a novel family of lipid phosphatases that dephosphorylate phosphatidic acid (PA) to produce diacylglycerol (DAG), and perform key functions in phospholipid and triacylglycerol biosynthesis and gene expression. In addition to its role in lipid biosynthesis, the yeast lipin Pah1p and its regulators are required for the maintenance of a spherical nuclear shape. This review summarizes recent advances in our understanding of the yeast lipin Pah1p and highlights the possible roles of phospholipid metabolism in nuclear membrane biogenesis.     

Microbial Relatives of Seed Storage Proteins: Conservation of Motifs in a Functionally Diverse Superfamily of Enzymes

Plant storage proteins comprise a major part of the human diet. Sequence analysis has revealed that these proteins probably share a common ancestor with a fungal oxalate decarboxylase and/or related bacterial genes. Additionally, all these proteins share a central core sequence with several other functionally diverse enzymes and binding proteins, many of which are associated with synthesis of the extracellular matrix during sporulation/encystment. A possible prokaryotic relative of this sequence is a bacterial protein (SASP) known to bind to DNA and thereby protect spores from extreme environmental conditions. This ability to maintain cell viability during periods of dehydration in spores and seeds may relate to absolute conservation of residues involved in structure determination. Received: 25 April 1997 / Accepted: 29 July 1997     

利用油体表达系统生产外源重组蛋白的研究进展

植物生产外源蛋白日益受到重视,是一个安全廉价的生产系统。植物油体表达系统利用油素蛋白的高表达性和易分离特性改进了植物生物反应器下游加工技术、降低了高纯度药用蛋白的生产成本。本文介绍了油体和油素蛋白的组成结构等特征,重点阐述了国内外各领域用植物油体表达系统生产外源蛋白的研究进展,探讨了油体系统的优势和存在的问题。本实验室利用油体系统开发酸性成纤维细胞生长因子(haFGF)医类新药,生物活性检测正在进行中。油体表达系统作为药用蛋白的新来源,将得到逐步的完善及更广泛的应用。     

Seed Lipids of the Grain Amaranths

The lipid and fatty acid composition of six members of two closelyrelated genera of the Amaranthaceae have been studied usinga combination of chromatographic procedures. The total lipidsvaried between 16.95 and 9.75% dry weight. Of these amounts,the non-polar lipids (mainly triglycerides) represented approximately90%, while polar lipids (glycolipids and phospholipids) accountedfor the remaining percentage. A relatively high degree of unsaturation was found for all lipidsdue to the dominance of linoleic acid (about 50%). The storage,metabolic, structural, and nutritional functions of the differentacyllipids are discussed.     

Membrane Lipids and the Conformations of Membrane Proteins

The general relations between protein conformation and the optical activity of peptide chromophores are outlined and applied to the analysis of the optical rotatory dispersion and circular dichroism of the plasma membranes of human erythrocytes and Ehrlich ascites carcinoma cells. It is concluded that the proteins of these membranes are "globular" and that they have considerable helical content. The spectroscopic consequences of perturbing the membranes with phospholipase C, phospholipase A, lysolecithin, and sodium dodecyl sulfate are examined in the light of the effects of these agents upon certain enzymatic and physical properties of the membranes and upon their proton magnetic resonance spectra. The data suggest that the architecture of membrane proteins is strongly dependent upon apolar lipid-protein and/or lipid-sensitive protein-protein interactions.     

An Assessment of Oxidative Damage to Proteins, Lipids, and DNA in Brain from Patients with Alzheimer's Disease

Abstract: Oxidative stress may contribute to neuronal loss in Alzheimer's disease (AD). The present study compares the levels of oxidative damage to proteins, lipids, and DNA bases from seven different brain areas of AD and matched control tissues by using a range of techniques. No differences in levels of lipid peroxidation were found in any of the brain regions by using two different assay systems. Overall, there was a trend for protein carbonyl levels to be increased in AD in frontal, occipital, parietal, and temporal lobe, middle temporal gyrus, and hippocampus, but a significant difference was found only in the parietal lobe. Gas chromatography-mass spectrometry was used to measure products of damage to all four DNA bases. Increased levels of some (8-hydroxyadenine, 8-hydroxyguanine, thymine glycol, Fapy-guanine, 5-hydroxyuracil, and Fapy-adenine), but not all, oxidized DNA bases were observed in parietal, temporal, occipital, and frontal lobe, superior temporal gyrus, and hippocampus. The baseline level of oxidative DNA damage in the temporal lobe was higher than in other brain regions in both control and AD brain. The finding of increased oxidative damage to protein and DNA strengthens the possibility that oxidative damage may play a role in the pathogenesis of AD in at least some key brain regions.