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91.
Jan Jonek Janusz Konecki Henryk Grzybek Zbigniew Olkowski 《Histochemistry and cell biology》1970,23(2):116-119
Summary Motoneurons in the spinal cords of exercised mice were investigated for acid phosphatase and thiamine pyrophosphatase by the electron microscope. In the control animals the acid phosphatase has been localized in lysosomes and in elongated vacuolar structures of the Golgi apparatus. The TPP-ase appears in vacuoles, vesicles and flattened cisternae of the Golgi apparatus.In conformity with a time of swimming the increase of TPP-ase and acid phosphatase activities was observed in the motoneurons of exercised animals. The authors suppose that lysosomes in the motoneurons of the exercised animals are formed in the Golgi zone, which enlarges according to the time of swimming. 相似文献
92.
Purification and Characterization of Myosin from Calf Brain 总被引:4,自引:1,他引:3
Mazhar N. Malik Michael D. Fenko Lois Scotto Pat Merz Jeffrey Rothman Helen Tuzio Henryk M. Wisniewski 《Journal of neurochemistry》1983,40(6):1620-1629
Actomyosin complex was extracted from the brain cortex in a medium consisting of low salt, ATP, and EDTA, in the presence of protease inhibitors, followed by ammonium sulfate fractionation. Myosin was then purified from the actomyosin. Myosin obtained according to the procedure used was significantly contaminated with actin high (greater than 200,000 dalton) and low molecular weight proteins. Therefore, an alternative method based on affinity chromatography (Blue Dextran/Sepharose) and gel filtration (Sepharose 4B) was developed to purify myosin. This procedure yielded myosin that was greater than 95% pure as judged by electron microscopy and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The subunit composition of purified brain myosin was monitored by sodium dodecyl sulfate-polyacrylamide gel also containing a urea gradient. A closely migrating triplet in the heavy chain and three light chains, LC1, LC2, and LC3, of Mr 21,000, 19,000, and 17,000, respectively, were observed. These findings raise the possibility of the existence of myosin isoenzymes in the brain. Brain myosin formed bipolar thick filaments in 0.075 M KCl and MgCl2. At low ionic strength, the Mg2+-ATPase activity of myosin was stimulated 3- to 3.5-fold in the presence of skeletal muscle f-actin. Brain myosin also hydrolyzed other nucleotides; the rate of hydrolysis was ITP greater than ATP approximately equal to CTP greater than GTP approximately equal to UTP. The substrate (ATP) saturation curve in the presence of 10 mM CaCl2 and 0.6 M KCl was complex and consisted of plateau regions. The Arrhenius plot of the Ca-ATPase data was linear, whereas with ITPase, it was biphasic with a break occurring around 20 degrees C. 相似文献
93.
We have studied the hydrodynamic properties of the complexes formed by interaction of nucleosome core particles with excess histone octamers containing two each of the four core histones. The results are consistent with tight binding of two to three octamers to the exterior of each core particle. The binding is dependent upon the presence of the H3/H4 histone pair: when H3/H4 alone are added to nucleosome core particles, tight binding is observed, but H2A/H2B alone are bound only weakly. We have also examined the properties of the nucleosome core in solutions containing 0·1 m to 0·7 M-NaCl. We show that in this salt range the core particle undergoes some changes in shape, reflected in a 14% increase in the frictional coefficient. Even at the highest salt concentrations used, however, the nucleosome core is still a compact, folded structure. 相似文献
94.
A retinol-binding protein (RBP), synthesized and secreted by ovine allantois in vitro, was purified from culture medium. The protein consisted of three isoelectric variants (pI 5.3-6.1) of identical molecular masses of about 23,000 Da as determined by two-dimensional PAGE under reducing conditions. Thirty-one of the first 34 N-terminal amino acids of the purified protein were sequenced and shown to have complete homology with bovine placental and bovine plasma RBP. The ultraviolet absorption spectrum and fluorescence excitation and emission spectra of the purified ovine placental RBP indicated the presence of bound retinol. Metabolic labeling studies demonstrated that the protein was synthesized by placental membranes. Using antiserum to bovine placental RBP, ovine placental RBP was immunolocalized in trophectoderm of 13-day-old blastocysts and trophectodermal cells of the chorion, endodermal cells lining the allantois, and ectodermal cells lining the amnion of 23-, 45-, and 53-day-old conceptuses. Results from this study suggest that ovine placental membrane epithelia synthesize and secrete RBP. Transport, storage, and metabolism of retinol mediated by placental RBP may be essential for normal embryonic development during pregnancy. 相似文献
95.
George A. Baumbach Philippa T. K. Saunders Catherine M. Ketcham Fuller W. Bazer R. Michael Roberts 《Molecular and cellular biochemistry》1991,105(2):107-117
Mature uteroferrin (Uf; M = 35,500) is a progesterone-induced acid phosphatase secreted by the pig uterus. It contains a single, unphosphorylated, high mannose-type oligosaccharide. Endometrial explants cultured in vitro secrete Uf with a M of 37,000 (37k Uf) having phosphorylated high mannose oligosaccharides. In this report we demonstrate that 37k Uf contains two N-linked oligosaccharides which are a mixture of complex and high mannose-type oligosaccharides. The complex-type glycopeptides are biantennary and a portion may be fucosylated on the GlcNac of the chitobiose core proximal to the peptide. Only a portion of the high mannose-type oligosaccharides are phosphorylated. The remainder appear to be typical Man6-4GlcNac2 oligosaccharides found on mature Uf.Abbreviations Uf
Uteroferrin
- ConA
Concanavalin A
- WGA
Wheat Germ Agglutinin
- endoH
endo--N-acetylglucosaminidase H
- SDS
Sodium Dodecyl Sulfate
- SDS-PAGE
polyacrylamide gel electrophoresis in the presence of SDS 相似文献
96.
G A Baumbach A H Climer N G Bartley H G Kattesh J D Godkin 《Biology of reproduction》1988,39(5):1171-1182
The major basic protein (BP) synthesized and secreted by elongating pig blastocysts was purified from medium of Day 14-17 conceptus cultures. Sequential ion-exchange and gel-filtration chromatographies resulted in isolation of BP as a single polypeptide of Mr = 43,100 or 42,800 under denaturing or native conditions, respectively. BP was found to be a glycoprotein by incorporation of [3H] glucosamine and susceptibility to N-glycopeptidase F. Two BP polypeptides were produced by N-glycopeptidase F (Mr = 39,800 and 36,300). Antiserum to BP immunoprecipitated radiolabeled BP from blastocyst culture medium. BP was not detected in medium from 1-2 mm diameter spherical (Day 10) blastocysts but was found in medium from 3-5 mm spherical (Day 10) and filamentous (less than 50 cm, Day 12) conceptuses, suggesting that BP synthesis and secretion began at the initiation of trophoblast expansion. With immunocytochemical procedures, BP was located in the apical cytoplasm of trophectoderm cells of Day 11 expanding (5-7 and 10-20 mm) blastocysts. These results suggest that trophoblast epithelium secrete BP apically toward the uterine lumen and that BP may play a role in maternal-fetal interactions during the peri-implantation period. 相似文献
97.
Time-resolved electron paramagnetic resonance (EPR) studies were carried out at room temperature and at 273 K on whole-cell samples of the photosynthetic algae: Anacystis nidulans and Scenedesmus obliquus, the latter being 97% deuterated from the growing medium. These photosynthetic organisms show greatly enhanced EPR signals which result from the generation of nonequilibrium spin populations, a phenomenon known as chemically induced dynamic electron polarization (CIDEP). We report magnetic-field profiles of the early transient signals of Photosystem I which are very similar to those observed at low temperatures. The results suggest that one or more early reduced electron acceptors in Photosystem I are being observed at ambient physiological temperatures. 相似文献
98.
H Sadeghi B S Wang A L Lumanglas J S Logan W R Baumbach 《Molecular endocrinology (Baltimore, Md.)》1990,4(12):1799-1805
GH specifically interacts with a soluble binding protein in serum. The GH-binding protein (GHBP) has been shown to contain the extracellular portion of the cell surface GH receptor (GHR). In rats and mice there is a unique mRNA that encodes the GHBP. This mRNA contains an alternatively spliced exon that replaces the transmembrane and intracellular domains of the receptor with a short hydrophilic carboxy-terminus of 17 and 25 amino acids, respectively, in rats and mice. In humans and other species no mRNAs encoding the GHBP have been identified, suggesting that the GHBP is in these cases a proteolytically processed GHR. In this study a monoclonal antibody (GHBP 4.3) was raised to the rat GHBP using as immunogen a synthetic peptide containing the unique C-terminal 17 amino acids that are not found in the rat GHR. As predicted, this antibody is specific to rat GHBP and does not cross-react with rat GHR. In combination with polyclonal and monoclonal antibodies that recognize both GHBP and GHR, this antibody was used to show that all, or most, of the GHBP in rat serum is indeed derived from the alternatively spliced GHBP mRNA and not from proteolytic processing of the GHR. In addition, endogenous rat serum GHBP was found to exist in two forms, with apparent mol wt of 52 and 44 kDa, arising from a single protein core of 32 kDa by extensive glycosylation. The concentrations of GHBP in male and female rat plasma were also estimated to be 300 and 575 ng/ml, respectively (measured in nonglycosylated GHBP equivalents). 相似文献
99.
Elizabeth J. White Henryk J. Juchniewicz John B. Clark 《Journal of neurochemistry》1989,52(1):154-161
Synaptosomes exposed to anoxic insult produce lactate at a slow rate (measured over 60 min). No measurable damaging effects were produced by prolonged depolarisation, anoxic insult, or exogenous lactate (2-32 mM) either on the synaptic plasma membrane (as judged by release of lactate dehydrogenase and soluble proteins), or on synaptosomal phospholipases (as judged by choline release from membrane phospholipids). Potassium-stimulated acetylcholine release was decreased by incubation in the presence of lactate (2-32 mM), as was potassium- and veratrine-stimulated calcium uptake and the calcium content of depolarised synaptosomes. The intrasynaptosomal pH was also reduced but there was no stimulation of oxygen radical production (as judged by H2O2 generation) by exogenous lactate. The role that lactic acidosis may play in giving rise to the altered calcium homeostasis and decreased acetylcholine release from synaptosomes exposed to anoxic insult is discussed. 相似文献
100.
Chmielewska A Konieczna L Plenis A Lamparczyk H 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2006,839(1-2):102-111
Reversed-phase-high-performance liquid chromatographic method with electrochemical detection has proven to be a highly sensitive and selective method for determination of trace components in complex biological samples, and the electrochemical detector becomes an important alternative tool to ultraviolet and fluorescence detectors. A rapid and sensitive method for the accurate determination of metoclopramide, hydrochlorothiazide, imipramine and diclofenac in serum or plasma samples is described. The method is based on liquid-liquid extraction. The compounds were separated on C-18 column as stationary phase with a different binary mixture as mobile phase. Proposed method was validated with respect to specificity, linearity range, limit of detection and quantitation, precision, accuracy and successfully applied in a pharmacokinetic studies. 相似文献