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1.
Recent studies have implicated accelerated sarcolemmal phospholipid catabolism as a mediator of the lethal sequelae of atherosclerotic heart disease. We have demonstrated that plasmalogens are the predominant phospholipid constituents of canine myocardium and that plasmalogens are hydrolyzed by a novel calcium independent plasmalogen selective phospholipase A2. Since the activities of phospholipases are modulated by the molecular dynamics and interfacial characteristics of their phospholipid substrates, we compared the molecular dynamics of plasmenylcholine and phosphatidylcholine vesicles by electron spin resonance spectroscopy and deuterium magnetic resonance spectroscopy. Plasmenylcholine vesicles have separate and distinct molecular dynamics in comparisons to their phosphatidylcholine counterparts as ascertained by substantial decreases in the angular fluctuations and motional velocities of probes attached to their sn-2 aliphatic constituents. Furthermore, since free radical oxidation of myocardial lipid constituents occurs during myocardial ischemia and reperfusion, we demonstrated that 1O2 mediated oxidation of plasmenylcholine resulted in the generation of several products which have chromatographic characteristics and molecular masses corresponding to 2-acyl lysophosphatide derivatives. Taken together, these studies underscore the biologic significance of the predominance of sarcolemmal plasmalogens present in mammalian myocardium and suggest that their catabolism by plasmalogen selective phospholipases and/or oxidative processes may contribute to the lethal sequelae of myocardial ischemia. 相似文献
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Using immunoblotting and immunofluorescent microscopy, we showed the presence in Xenopus laevis oocytes of two prosomal proteins (27 and 31-33 kDa) and studied their distribution during oogenesis. In the ooplasm, both proteins are detected in prosomal clusters of various size. During previtellogenesis, prosomal proteins are diffusely distributed in the nucleoplasm and form evenly distributed clusters in the cytoplasm. During oocyte growth, prosomal proteins disappear from the nucleus and form animal-vegetal and cortical gradients in the cytoplasm. In the course of oocyte maturation prosomal clusters become smaller. After artificial activation of the egg, the dorso-ventral gradient of distribution of prosomal proteins is observed. Double immunohistochemical labeling revealed morphological association between prosomal clusters and fibril-like structures of the oocyte containing actin and myosin. The latter are then replaced by diffusely distributed actin and myosin. Thus, correlation is observed between localization of the acto-myosin complex of the oocyte and that of prosomal proteins. 相似文献
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Prosomes were first described as being mRNA-associated RNP (ribonucleoprotein) particles and subcomponents of repressed mRNPs (messenger ribonucleoprotein). We show here that prosomes isolated from translationally inactive mRNP have a protease activity identical to that described by others for the multicatalytic proteinase complex (MCP, 'proteasome'). By RNase or non-ionic detergent treatment, the MCP activity associated with repressed non-globin mRNP from avian erythroblasts, sedimenting at 35 S, could be quantitatively shifted on sucrose gradients to the 19-S sedimentation zone characteristic of prosomes, which were identified by monoclonal antibodies. The presence of small RNA in the enzymatic complex was shown by immunoprecipitation of the protease activity out of dissociated mRNP using a mixture of anti-prosome monoclonal antibodies; a set of small RNAs 80-120 nucleotides long was isolated from the immunoprecipitate. Furthermore, on CsCl gradients, colocalisation of the MCP activity with prosomal proteins and prosomal RNA was found, and no difference in the prosomal RNA pattern was observed whether the particles were fixed or not prior to centrifugation. These data indicate that the MCP activity is a property of prosomes, shown to be in part RNP and subcomplexes of in vivo untranslated mRNP. A hypothesis for the role of the prosome-MCP particles in maintaining homeostasis of specific protein levels is proposed. 相似文献
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Structural and functional reconstitution of the glucocorticoid receptor-hsp90 complex 总被引:11,自引:0,他引:11
L C Scherrer F C Dalman E Massa S Meshinchi W B Pratt 《The Journal of biological chemistry》1990,265(35):21397-21400
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Tatsuo Iwasa Kazuo Takeda Fumio Tokunaga Peter S. Scherrer Lester Packer 《Bioscience reports》1982,2(11):949-958
To elucidate the role of tyrosine residues in the shift of max and the light-driven proton pump of bacteriorhodopsin~ the photochemical reaction of tyrosine-iodinated bacteriorhodopsin (tyr-mod-bR) was investigated by low-temperature spectrophotometry. After 4–5 of 11 tyrosine residues of bacteriorhodopsin were iodinated, the meta-intermediate of tyr-mod-bR in 75% glycerol solution became so stable that its decay could be observed even at room temperature and i t was stable in the dark for several hours at –65°C.Four batho-intermediates were formed by irradiation with green light (500 nm) at –170°C. Like native bacteriorhodopsin, these batho-intermediates were photoreversible at –170°C. Four corresponding meta-intermediates were also formed by irradiation at –60°C. Using the difference spectra between meta-intermediates and tyr-mod-bR, the absorption spectra of four kinds of tyr-mod-bRs, batho-intermediates, and meta-intermediates were estimated. Each was at shorter wavelengths than that of its corresponding type in native bacteriorhodopsin. The results indicate that two or more tyrosine residues have some role in determining color in native bacteriorhodopsin. 相似文献
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A rapid and sensitive method for detection of proteins in polyacrylamide SDS gels: Staining with ethidium bromide 总被引:4,自引:0,他引:4
We describe here a fluorometric method of detection of proteins fractionated by electrophoresis in polyacrylamide-SDS gels. This method, using ethidium bromide as fluorescent dye, is performed within 40 minutes after the end of the electrophoretic run. It does not require treatment of proteins prior to electrophoresis, and entails neither fixation of proteins in the gel, nor destaining. It is sufficiently sensitive to detect 0.5–1.0 g of protein per band. Furthermore, the simultaneous electrophoretic resolution and detection of protein and RNA on a single SDS-polyacrylamide gradient gel is reported. 相似文献
10.
Isolation and partial characterization of a biosynthetic N-terminal methionyl peptide of bovine pars intermedia: relationship to ubiquitin 总被引:1,自引:0,他引:1
N G Seidah P Crine S Benjannet H Scherrer M Chrétien 《Biochemical and biophysical research communications》1978,80(3):600-608
During in vitro labelling studies of beef pituitary intermediate lobe cells, a 4000 daltons molecular weight peptide was found to be biosynthesized in major yields. The partial amino acid sequence of this peptide has been found to be Met 1, Leu 8, 15 and Lys 6, 11, 27, 29, 33. This partial sequence fits very well the one expected from the N-terminal sequence of bovine and human ubiquitin, a non-histone fragment of the nuclear protein A24. Since an identical peptide was also biosynthesized from rat hypothalamus and mouse AtT-20 tumor cells, the ubiquitous nature of this peptide is further revealed. 相似文献