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11.
Binding of BiP to an assembly-defective protein in plant cells 总被引:5,自引:1,他引:4
Emanuela Pedrazzini Giovanna Giovinazzo Roberto Bollini Aldo Ceriotti Alessandro Vitale 《The Plant journal : for cell and molecular biology》1994,5(1):103-110
The binding protein (BiP) has been implicated as a mediator of protein folding and assembly in the endoplasmic reticulum of mammalian cells and has often been found in stable association with structurally defective proteins. To acquire information on the activity of BiP in plant cells, we have expressed in tobacco protoplasts the wild type form and an assembly-defective form of bean phaseolin. Phaseolin (PHSL) is a soluble, trimeric, storage glycoprotein co-translationally inserted into the lumen of the endoplasmic reticulum and then transported along the secretory pathway to the protein storage vacuoles. We have previously shown that a PHSL mutant in which the last 59 amino acids have been deleted (Δ363PHSL) is unable to form trimers and is retained in a pre-Golgi compartment when synthesized in Xenopus oocytes. When transiently expressed in tobacco leaf protoplasts, wild-type PHSL is correctly glycosylated and assembles efficiently and rapidly into trimers. Δ363PHSL is also correctly glycosylated but does not trimerize. Tobacco BiP and Δ363PHSL are co-immunoselected using either anti-PHSL or anti-BiP antibodies. Under the same conditions, co-immunoselection of BiP with wild-type PHSL is not detectable. The BiP bound to Δ363PHSL can be released by treatment of the complex with ATP, indicating that the binding is related to the proposed function of BiP in protein folding and assembly in the endoplasmic reticulum. These data indicate that BiP stably binds structurally defective proteins in plant cells. 相似文献
12.
The role of rapidly exchanging intracellular Ca2+ stores in the control of Ca2+ homeostasis is reviewed. The following issues are discussed: the reasons why such stores exist in eukaryotic cells; the differences between the terminal cisternae of the skeletal muscle sarcoplasmic reticulum, which have direct, physical connection with the T tubules of the plasmalemma, and the Ca2+ stores located in the depth of the cytoplasm, which are stimulated by second messengers; the cytological nature (subcompartments of the ER) of the rapidly exchanging Ca2+ stores and their functional significance. The conclusions introduce recent developments in which intracellular Ca2+ stores have been investigated also by molecular biology techniques. 相似文献
13.
Arturo A. Vitale Carina C. Ferrari Hernán Aldana Jorge M. Affanni 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1996,681(2):381
In the present study a new chromatographic method was developed to quantify melatonin in rat pineal that can be extended to other tissues. Melatonin was extracted from an acid homogenate with ethyl acetate to avoid amine interference. HPLC was performed with silica normal-phase column and fluorescence detection. This method is sensitive enough for detecting melatonin in a single pineal gland with a detection limit of 3 pg/mg tissue. 相似文献
14.
From filtrates of an oxytetracycline-producing culture of Streptomyces rimosus a deoxyribonuclease was purified to homogeneity and determined to be a potent endo-DNase. It is a monomeric, basic protein (Mr 21 000; pI 9.5) stable in a broad pH range but unstable to higher temperature. The enzyme has an absolute requirement for Mg2 + or Mn2 +, and for its full activity requires free SH groups and a low-ionic-strength environment. Its N-terminal primary structure differs from that of other nucleases. 相似文献
15.
Gene Expression and Synthesis of Phytohemagglutinin in the Embryonic Axes of Developing Phaseolus vulgaris Seeds 总被引:2,自引:1,他引:1 下载免费PDF全文
Phytohemagglutinin (PHA), the major seed lectin of the common bean (Phaseolus vulgaris), is found largely in the cotyledons, but is also present in the embryonic axis. At mid-maturation, the percentage of total protein synthesis which is directed towards making PHA is 5 to 10 times greater in the cotyledons than in the axes. This lower rate of synthesis in the axes is correlated with a lower abundance of mRNA for PHA, as determined by dot blot hybridization using a cDNA clone for PHA. Manen and Pusztai (Planta 1982 155: 328-334) have claimed on the basis of immunocytochemical evidence that, in the axis, PHA is found in the cytosol although it is present in protein bodies in the cotyledons. In the cotyledons, PHA is synthesized on rough endoplasmic reticulum, and its transport to the protein bodies via the Golgi complex is associated with specific posttranslational processing steps (Vitale and Chrispeels, J Cell Biol 1984 In press). A cytosolic localization of axis PHA would be an indication of a different site of synthesis and transport pathway. The results presented here indicate that the site of synthesis of PHA and the posttranslational modifications of PHA are the same in the axes as in the cotyledons. Since in the cotyledons these modifications take place in the endoplasmic reticulum, the Golgi, and the protein bodies, it appears that the transport pathway and the site of accumulation of PHA in the axes is similar to that in the cotyledons. On the basis of our evidence, we suggest that the subcellular localization of PHA in the axes should be reexamined. 相似文献
16.
Phytohemagglutinin, the major lectin in the seeds of the common bean Phaseolus vulgaris L., was isolated by affinity chromatography from cotyledons of nearly mature seeds and from developing cotyledons labeled with [3H]glucosamine, [3H]mannose or [3H]fucose. The protein was subjected to exhaustive proteolysis and the carbohydrate composition of the resulting glycopeptides examined. Two classes of oligosaccharide side-chains were found. The sidechains of the first class are of the high-mannose type, containing two residues of N-acetylglucosamine and 8 or 9 mannose residues. The sidechains of the second class are of the modified type containing N-acetylglucosamine, mannose, fucose, xylose in molar ratios of 2:3.8:0.6:0.5. Two-dimensional gel electrophoresis shows that phytohemagglutinin can be fractionated into seven different glycosylated polypeptides, and that each one contains at least one modified oligosaccharide chain. The results indicate that most glycosylated polypeptides probably contain one chain of each class. The carbohydrate composition of the two types of chains is similar to that found in other plant glycoproteins, but this is the first report of a plant glycoprotein with both highmannose and modified oligosaccharides on the same polypeptide chain.Abbreviations endo H
endo--N-acetylglucosaminidase H
- GlcN
glucosamine
- GlcNAc
N-acetylglucosamine
- Man
mannose
- PHA
phytohemagglutinin
This work was done while A.V. was on leave from the Istituto Biosintesi Vegetali, C.N.R., via Bassini 15, I-20133 Milano, Italy 相似文献
17.
Blattella germanica (L.) that were fed hydramethylnon bait produced residues that were toxic to exposed conspecifics. Insecticidal activity was traced to the feces of treated insects by feeding radiolabeled material, where approximately 50% of the recovered radioactivity was unmetabolized parent compound. Ingestion of toxicant-laden feces by all life stages was evident, but the effect of this behavior was greatest on early instar nymphs. Baits containing toxicants with delayed activity, such as hydramethylnon, probably affect cockroach field populations indirectly through coprophagy. 相似文献
18.
19.
Dr. Darrell R. Stokes Anthony J. Vitale Charles R. Morgan 《Cell and tissue research》1979,198(1):175-189
Histochemical techniques have been employed to characterize enzymatic activity in the mesocoxal muscles of the cockroach, Periplaneta americana. Through our studies of the enzymes myosin-ATPase, NADH reductase, succinic dehydrogenase (SDH), and lactic dehydrogenase (LDH), we were able to classify fibers within these muscles according to criteria established for muscle fibers of vertebrates. Many of the mesocoxal muscles possess two different and distinct populations of fibers, whereas the remaining muscles are homogeneous with respect to their constituent fibers. The data presented here indicate biochemical heterogeneity for muscles of differing structural and functional features and possible neurotrophic influences upon oxidative enzymes and myosin-ATPase isozymes. 相似文献
20.