Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0272170 (SDS)
 50,377 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We investigated the allergenicity of pollen extract of Japanese juniper (Juniperus rigida, Cupressaceae family) to sera of 49 asthmatic children by the methods of enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Three bands stained with Coomassie Brilliant Blue R were detected on SDS-PAGE. Sera from 27 (55.1%) out of the 49 children showed positive reaction to the pollen extract in ELISA. The same sera from the 27 children were used as the first antibody in immunoblotting, which confirmed the presence of a band of protein with 70 K dalton molecular weight (M.W) common to the all sera. This band was bound with concanavalin A in lectins. We successfully purified the antigenic substance of Japanese juniper pollen from this band by the electroelution method. The major allergen of Japanese juniper pollen is glycoprotein with 70 KM.W. On sandwich-ELISA, there was no reaction of Sugi Basic Protein (SBP) and anti-SBP to Japanese juniper is an allergen that has no cross-reactivity with SBP.
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PMID:[Evaluation of allergenicity of the extract of Japanese juniper pollen reacting to sera from asthmatic children--by the methods of enzyme-linked immunosorbent assay and immunoblotting]. 148 95

The 5SrRNA in the rat liver postmicrosomal supernatant was investigated. Acrylamide gel electrophoresis and Northern blot analysis showed that most of the 5SrRNA was present in the fractions obtained on high molecular weight regions separated by Sephadex G-200 column chromatography of the supernatant, which contained the bulk of the methionyl-tRNA synthetase (Fraction I) and tyrosyl-tRNA synthetase (Fraction II). A high molecular weight complex containing nine aminoacyl-tRNA synthetases [Mirande, M., LeCorre, D., & Waller, J.-P. (1985) Eur. J. Biochem. 147, 281-289] was purified by fractional precipitation with polyethylene glycol 6000, gel filtration on Bio-Gel A-1.5m, and finally tRNA-Sepharose column chromatography, which gave two fractions. Fraction B showed the activities of nine aminoacyl-tRNA synthetases and gave protein bands corresponding to eight previously identified enzymes on SDS-PAGE. Fraction A, eluted with a lower KCl concentration than Fraction B, showed lower activities than fraction B of eight of the aminoacyl-tRNA synthetases, the exception being prolyl-tRNA synthetase. The staining patterns with ethidium bromide of the RNAs after PAGE showed 5SrRNA bands for Fraction A but not for Fraction B. However, Northern blot analysis indicated that 5SrRNA was present in both Fractions A and B. The staining pattern after SDS-PAGE of Fraction A with Coomassie Brilliant Blue showed several protein bands in addition to those observed for Fraction B, one of which, with a staining intensity comparable with those of other bands, was located at the same position as ribosomal protein L5, which is the protein moiety of the 5SrRNA-L5 protein complex of ribosomal 60S subunits.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Occurrence of 5SrRNA in high molecular weight complexes of aminoacyl-tRNA synthetases in a rat liver supernatant. 179 73

Alkaline phosphatase (ALP) activity has been demonstrated in periodontal ligament (PDL). On the basis of electron microscopic study, distribution of the enzyme in PDL tissue has also been indicated not only as a cell associated activity but also as an extracellular matrix associated activity. This study is concerned with the purification and characterization of the enzyme obtained from bovine PDL tissue. Purification of ALP extracted from the tissue included solubilization with 10 mM Tris-HCl buffer, pH 7.4, containing 0.2 mM MgCl2 and 0.1% Nonidet P-40 and fractionation by sequential chromatography utilizing DEAE-sephacel, Sepharose CL-6B and concanavalin A Sepharose 4B. Purity was established by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). This was followed by staining for ALP activity first with 2 mM beta-naphthyl acid phosphate and 1 mM Fast Blue BB Salt and then the protein with Coomassie Brilliant Blue. SDS-PAGE of the crude enzyme preparations gave a broad band with apparent molecular weight of 110,000-130,000 dalton. ALP activities were separated into two major peaks using Sepharose CL-6B chromatography. The void volume peak showed a purified form of 110,000 dalton ALP (110K ALP) while the second peak contained 120,000-130,000 dalton ALP (120-130K ALP) and other proteins. Sequentially, 120-130K ALP was purified by chromatography on concanavalin A Sepharose 4B. A polyclonal antibody was raised against purified bovine PDL 110K ALP in a rabbit. Immunodiffusion analysis showed that a polyclonal antibody against 110K ALP recognized 120-130K ALP. Analytical affinity chromatography on concanavalin A Sepharose 4B indicated that 110K ALP and 120-130K ALP had distinct affinity to the column which may depend upon the sugar chain structure. Digestion of 110K ALP with phosphatidylinositol-specific phospholipase C affected electrophoretic mobility but 120-130K ALP had no effect. It is suggested that 110K ALP is attached to a cell membrane anchored by a phosphatidylinositol glycan. In conclusion, bovine PDL contains two types of alkaline phosphatase i.e. 110K ALP and 120-130K ALP. Both ALPs are immunologically related although they have different sugar chain moieties. Furthermore, 110K ALP has a membrane anchoring domain. These results suggest that 110K ALP would be localized on the surface of the cell membrane and 120-130K ALP may associated with the extracellular matrix.
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PMID:[Purification and characterization of alkaline phosphatase obtained from bovine periodontal ligament]. 213 40

An antigen was isolated and partially characterized from adult Schistosoma mansoni which immunologically cross-reacted with Fasciola hepatica and Biomphalaria glabrata, a schistosome intermediate snail host. The antigen was isolated from a solubilized freeze-thaw preparation containing 0.5% Triton X-100 by passage through a CNBr-activated Sepharose 4B column coupled with rabbit IgG prepared against a homogenate of B. glabrata hepatopancreas. The eluted antigen (designated as SMw-53P) stained with Coomassie Brilliant Blue R250, but not with Nile Blue A or periodic acid-Schiff's stains. The antigen did not bind to a Concanavalin A affinity column. SMw-53P was determined to have a molecular weight of 53,000 daltons by SDS-polyacrylamide gel electrophoresis. Western blotting, using sera from mice infected with S. mansoni, revealed the presence of the antigen in whole worm preparations of both S. mansoni and F. hepatica. The serodiagnostic potential of the antigen was evaluated by ELISA utilizing sera from S. mansoni-infected mice, or rabbits injected with homogenates of F. hepatica or S. mansoni whole worm, or B. glabrata hepatopancreas. SMw-53P was shown to strongly react with all antisera, but not normal mouse or rabbit sera. These data suggest a limited value for the antigen for the specific immunodiagnosis of schistosomiasis mansoni, but do suggest a possible potential as a general screening tool for detecting trematode infections. Further studies regarding the antigen's potential as a vaccine are indicated.
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PMID:Isolation and partial characterization of an antigen shared between Schistosoma mansoni, Fasciola hepatica, and Biomphalaria glabrata. 241 36

Saito et al recently reported that the alkaline phosphatase (ALPase) of human periodontal ligament fibroblasts (HPLF) showed remarkably high activity which was similar, but not identical phenotype, to that present in osteoblasts, and suggested that HPLF could be termed as "osteoblastic fibroblast." The present study attempts to explore the ALPase synthesized on HPLF in relation to 1,25(OH)2D3. These HPLF were obtained by the explantation method and then subcultured in D-MEM containing 2 mg FCSP/ml, 50 micrograms ascorbic acid/ml and penicillin/streptomycin after trypsinization. The HPLF were inoculated at a cell density of 1.25 x 10(4) cells/cm2 in culture wells. After 24hr, the HPLF were treated every two days for 7 days with 0.5-10nM 1,25 (OH)2D3. Then, ALPase activity, DNA and protein contents were assayed by the methods using p-nitrophenylphosphate, diaminobenzoic acid and Coomassie Brilliant Blue, respectively. Also, ALPase was prepared from the confluent HPLF incubated with 5 nM 1,25 (OH)2D3 for 12 days, and digested with and without trypsin. The crude ALPase which was solubilized with 10mM Tris-HCl, pH 7.4 containing 0.2 mM MgCl2 and 0.1% NP-40 was applied to 5-15% gradient SDS-PAGE and stained with beta-naphththylacid phosphate and First Blue BB salt in 60 mM borate buffer pH 9.7. The cell growth which was assayed by DNA contents and the incorporation of 3H-thymidine was decreased by 1,25(OH)2D3. On the other hand, ALPase activity was increased approximately 3.6 fold at 6 day by the addition of 5 nM 1,25(OH)2D3. From the separation of ALPase activity on SDS-PAGE, 110 K and 120-130 K ALPase were identified. The 110 K ALPase, which was not changed by 1,25(OH)2D3, was converted to 100K, releasing 10K peptide after trypsin treatment. This 110K ALPase might be tightly associated with cell membrane structure. The 120-130K ALPase was remarkably increased by 1,25(OH)2D3 on SDS-PAGE and completely digested with trypsin. The ALPase in the cultured HPLF might be located not only on the plasma membrane but also in the extracellular matrix. Therefore, 1,25(OH)2D3 may regulate the cell cycle and also the gene expression of ALPase of HPLF.
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PMID:[Biochemical study of human periodontal ligament fibroblasts--1,25 (OH)2D3 dependent alkaline phosphatase]. 248 52

The purification of cytochrome P-450-dependent 14 alpha-sterol demethylase (P-450DM) from the important opportunistic fungal pathogen, Candida albicans, is described. Optimal purification (875-fold) was achieved by extracting the cytochrome from microsomes with sodium cholate followed by hydroxyapatite, octyl-Sepharose and CM-Sepharose chromatographies, giving a cytochrome preparation of 17.5 nmol/mg of protein. By the use of SDS/polyacrylamide-gel electrophoresis the cytochrome was judged to be highly purified on the basis of Coomassie Brilliant Blue staining of protein. The Mr of P-450DM was estimated to be 51,000. The absorption spectrum of oxidized P-450DM was characteristic of a low-spin cytochrome, and its reduced CO complex had a Soret absorption peak at 447 nm. When reconstituted in a model membrane system of dilauroylphosphatidylcholine with NADPH and O2, P-450DM catalysed the complete 14 alpha-demethylation of lanosterol, which was inhibited by CO. The cytochrome appeared to have a high degree of substrate specificity; it was unable to oxidize a number of xenobiotic compounds in the reconstituted assay.
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PMID:Purification and properties of cytochrome P-450-dependent 14 alpha-sterol demethylase from Candida albicans. 255 99

Strains of Saccharomyces cerevisiae expressing P28-I, an antigen inducing protection against schistosomiasis, have been constructed. Transformants containing a very high copy number of a P28-I expression vector were selected by genetic complementation involving deficient LEU2 or URA3 alleles carried by plasmids. Using the ura3 fur1 auto-selection system, constitutive and stable expression of P28-I could be obtained in cultures grown in rich medium. The accumulation of the foreign protein exceeds 25% of total yeast proteins when estimated by Coomassie Brilliant Blue staining of SDS-PAGE. Moreover, P28-I which was located intracellularly was soluble and biologically active.
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PMID:High level of expression of a protective antigen of schistosomes in Saccharomyces cerevisiae. 269 78

The human T cell leukemia virus type I (HTLV-I)-transformed T lymphocyte cell line MT-2 constitutively produces differentiation-inducing factor (DIF) for the human promyelocytic leukemia cell line HL-60. Purification and characterization of DIF derived from MT-2 were performed here to identify T cell-derived DIF. DIF was purified from conditioned medium of the MT-2 cell culture with serum-free medium by utilizing the sequential chromatographies of anion-exchange (mono-Q) column, gel filtration (superose-12) column, and hydrophobic (phenyl 5PW) column and finally the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). In the SDS-PAGE, one major band with a molecular weight of 56,000 and one minor band with 15,000 were stained with Coomassie Brilliant Blue and both showed DIF activity after extraction from gels. DIF had an isoelectric point of 5.8. In all purification steps, DIF activity for HL-60 and cytotoxic activity to the sublines of mouse L-929 were not able to be separated. Further, DIF was neutralized by antibody against lymphotoxin (LT) but not by antibody against tumor necrosis factor. These results indicate that the major DIF activity derived from MT-2 is LT.
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PMID:Purification and characterization of a T lymphocyte-derived differentiation-inducing factor for human promyelocytic cell line (HL-60) and its relationship to lymphotoxin. 278 94

Saliva samples were obtained from the cannulated duct of the rat submandibular gland following intravenous administration of the autonomic agonists, acetyl beta-methyl choline (methacholine), phenylephrine and isoprenaline and by stimulating sympathetically at 5 and 20 Hz. Phenylephrine and isoprenaline produced a saliva approximately 50-fold richer in protein than that induced by methacholine. Protease activity was greatest in saliva induced by phenylephrine. Methacholine, phenylephrine and isoprenaline caused the secretion of markedly different protein populations, determined by separation on SDS-polyacrylamide gradient gels followed by staining with Coomassie Brilliant Blue R250. Many species in the molecular weight range 10000-40000 were secreted in response to phenylephrine and isoprenaline. Methacholine produced a saliva poor in protein and the resulting electrophoretic banding patterns were faint. Staining with periodic acid-Schiff's reagent showed that the largest protein species (mol. wt. 150000) secreted in response to isoprenaline was glycosylated and that small quantities of a similar protein were present in saliva produced by methacholine and phenylephrine but were not visualized by staining with Coomassie Brilliant Blue R250. Phosphorylated proteins of molecular weights 30000-35000 and 38000-45000 were detected in saliva produced by isoprenaline but not in saliva produced by methacholine or phenylephrine. Stimulation of the cervical sympathetic trunk at 5 Hz yielded a population of salivary proteins, many of which behaved similarly on electrophoresis to proteins secreted in response to isoprenaline. At 20 Hz a different population was secreted, resembling that present in phenylephrine-induced saliva.
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PMID:The effect of autonomic agonists and nerve stimulation on protein secretion from the rat submandibular gland. 285 87

Rat liver coated vesicle preparations were frequently found to contain small ovoid bodies, which resembled coated vesicles in morphology. We have purified these bodies to homogeneity using sucrose density gradients and preparative agarose gel electrophoresis. When negatively stained and viewed by electron microscopy, the purified structures display a very distinct and complex morphology, resembling the multiple arches which form cathedral vaults. They measure 35 X 65 nm and are therefore considerably larger than ribosomes. When subjected to SDS PAGE, these structures, which we refer to as vaults, appear to contain several minor and five major species: Mr 210,000, 192,000, 104,000, 54,000, and 37,000. One of these (Mr 104,000) greatly predominates, accounting for greater than 70% of the total Coomassie Brilliant Blue-staining protein. Another major species of Mr 37,000 has been identified as a species of small RNA of unusual base composition (adenosine 12.0%, guanosine 29.7%, uridine 30.9%, and 27.4% cytidine), which migrates as a single species in urea PAGE between the 5S and 5.8S ribosomal standards, containing approximately 140 bases. Although the RNA constitutes only 4.6% of the entire structure, the large size of the particle requires that each one contains approximately 9 molecules of this RNA. Antibodies prepared against the entire particle are largely specific for the major (Mr 104,000) polypeptide species. Although they do not directly react with the RNA constituent on Western blots, these antibodies immunoprecipitate a 32P-labeled RNA of identical size from metabolically-labeled rat hepatoma cells. Vaults are observed in partially purified fractions from human fibroblasts, murine 3T3 cells, glial cells, and rabbit alveolar macrophages. It therefore appears that these novel ribonucleoprotein structures are broadly distributed among different cell types. The function of vaults is at present unknown.
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PMID:Isolation and characterization of a novel ribonucleoprotein particle: large structures contain a single species of small RNA. 294 44


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