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)

Neurofilaments were isolated from desheathed and minced segments of rat peripheral nerve by osmotic shock into 0.01 M Tris-HCI buffer, pH 7.2. Freshly isolated neurofilaments were observed to undergo disassembly by progressive fragmentation upon exposure of dilute tissue extracts to this buffer. Low- and high-speed centrifugations of these tissue extracts separated membranous and particulate constituents and produced a progressive enrichment of 68,000-dalton polypeptide band in successive supernates, as determined by analyses of soluble proteins by SDS-polyacrylamide electrophoresis. The final high-speed supernatant fractions (S3) of nerve extracts, which were predominantly composed of 68,000-dalton polypeptide, were used to raise a specific experimental antisera in rabbits. Utilizing techniques of immune electron microscopy, experimental rabbit antisear was shown to contain antibodies against neurofilaments. Intact neurofilaments isolated from rat nerves and attached to carbon-coated grids became decorated when exposed to experimental rabbit antisera or purified gamma globulin (IgG) derivatives. The decoration of neurofilaments closely resembled the IgG coating seen in immune electron microscopy. Antibody absorption techniques were used to identify the biochemical constituency of neurofilamentous antigenic determinants. The decoration of neurofilament by experimental IgG was not altered by additions of tubulin or bovine serum albumin, but was prevented by additions of S3 fractions as well as the 68,000-dalton polypeptide of this fraction which was eluted and recovered from polyacrylamide gels. These findings are indicative that a 68,000-dalton polypeptide is a constituent subunit of rat peripheral nerve neurofilaments.
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PMID:Immunological and ultrastructural studies of neurofilaments isolated from rat peripheral nerve. 6 60

Colony stimulating factor-1 (CSF-1) is a homodimeric glycoprotein that humorally regulates the proliferation and differentiation of mononuclear phagocytic cells and locally regulates cells of the female reproductive tract. Alternative splicing of the human CSF-1 mRNA leads to alternative expression of the CSF-1 homodimer as a secreted glycoprotein or as a membrane-spanning molecule with cell surface biological activity. In the present study, analysis of immunoaffinity-purified CSF-1 from mouse L929 cell medium by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) indicated that CSF-1 is predominantly secreted as highly sulfated species of 375- and 250-kDa with a smaller amount of a 100-kDa species. Analysis by gel filtration in 4 M guanidine HCI buffer, indicated that, in contrast to the 100-kDa species, the highly sulfated species exhibit anomalously high molecular weights and self-association on SDS-PAGE similar to the dermatan sulfate proteoglycan, biglycan. The three predominant CSF-1 species were shown to be an 80-kDa homodimer, an 80-kDa/50-kDa heterodimer, and a 50-kDa homodimer. The 80-kDa subunit contained a single 18-kDa chondroitin sulfate chain that was absent from the 50-kDa subunit. Furthermore, treatment of the 80- and 50-kDa subunits, synthesized in the presence of tunicamycin, with chondroitinase ABC, neuraminidase, and endo-alpha-N-acetyl galactosaminidase reduced their apparent molecular masses to 60 and 25 kDa, respectively. These results are consistent with intracellular proteolytic cleavage of the 80-kDa chondroitin sulfate containing subunits from the membrane spanning CSF-1 precursor at a point carboxyl-terminal to the single consensus sequence for glycosaminoglycan addition and cleavage of the 50-kDa glycoprotein subunit at a position aminoterminal to this site. The predominance of the proteoglycan form of secreted CSF-1, which represents only 3-4% of the total trichloroacetic acid-precipitable counts released from 35SO4(2-)-labeled L cells, has important implications for regulation by this growth factor.
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PMID:The predominant form of secreted colony stimulating factor-1 is a proteoglycan. 173 26

Proteins were extracted from rat liver ribosomes and ribosomal subunits: with 67% acetic acid (in the presence of 3.3 mM, 33 mM, or 67 mM Mg) with 2 M LiCL in 4 M urea; with 0.25 N HCI; with 1% SDS; and after RNase digestion. The most efficient extraction and the best recovery were either with acetic acid in the presence of 33 mM or 67 mM Mg, or with LiCI-urea. Protein extracted with acetic acid, LiCi-urea, or with HCI had little or no contamination with RNA. The ribosomal proteins were analyzed by two-dimensional polyacrylamide gel electrophoresis: the proteins extracted with acetic acid were the most soluble in the sample gel solution; their electrophoretograms displayed the maximum number of spots and the smallest number of derivatives or altered proteins. Preparations of protein extracted with SDS or RNase were relatively insoluble in the sample gel solution, and proteins extracted with HCI showed a large number of derivatives. All things considered, the most satisfactory method for the extraction of protein from eukaryotic ribosomes is with 67% acetic acid in the presence of 33 mM MgCl2.
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PMID:The extraction of proteins from eukaryotic ribosomes and ribosomal subunits. 445 58

Tumor rejection antigen (TRA) of an ultraviolet-light-induced murine skin tumor was solubilized, fractionated and partially characterized. Subcellular fractions were prepared by differential centrifugation of tumor cells that had been ruptured via nitrogen cavitation. Only the 110,000-g membrane fraction induced significant tumor protection, as determined by in vivo immunization and challenge assays. Extraction of the membrane fraction with 3 M KCI resulted in solubilization of material that could induce in vivo tumor-rejection immunity. Both the membrane fraction and soluble extract had a limited effective dose range. The KCI extract was separated on a Sepharose, CL-6B column in the presence of 6 M guanidine-HCI. Only one of five fraction pools (molecular weight range of 76,000-127,000 daltons) was immunogenic. It contained at least eight protein bands by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), but no lipid components. This immunogenic Sepharose fraction was chromatographed on a Sephadex G-150 column. Each of the four Sephadex fraction pools was immunogenic. One protein component was common to each of those fractions. It migrated as a single 76,000-dalton band on SDS-PAGE and contained [14C]-leucine and [3H]-glucosamine that had been incorporated during cell growth. These results suggest that the TRA of this tumor is a 76,000-dalton glycoprotein.
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PMID:Solubilization and partial characterization of a tumor-rejection antigen from an ultraviolet light-induced murine tumor. 727 57

Sesamoid bone cartilage from the metacarpophalangeal (MCP) joints of six-month-old calves was cultured intact on its bone support. The sesamoid bones for the experiment were subjected to loading with 0.2 MPa at 0.2 Hz for a week. Controls were cultured for 0 or 7 days, labelled with [35S] sulfate for 17 h, harvested, and extracted in 4 M guanidine HCI with proteinase inhibitors, for analysis on Sepharose CL-2B columns under dissociative conditions. It was found that the population of labelled small proteoglycans was significantly larger in the loaded cartilage than in the cultured controls. This population was pooled and further purified on dissociative Sepharose CL-4B columns. The resulting profiles showed two peaks. The material eluting at Kav 0.80 contained only chondroitin sulfate and keratan sulfate chains. The peak at Kav 0.48 was analysed by SDS-polyacrylamide gel electrophoresis. It consisted of the dermatan sulfate proteoglycans decorin and biglycan. The amounts of newly synthesized decorin and biglycan in the cultured control cartilage were lower than in the control cartilage that had been labelled on day 0. The synthesis of decorin, but not of biglycan, was significantly higher in the loaded cartilage than in the cultured control cartilage. Through its interaction with collagen type II, decorin may aid the adaptation of articular cartilage to increased loading.
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PMID:Mechanical loading affects the synthesis of decorin and biglycan in intact immature articular cartilage in vitro. 755 63

An immunoaffinity column was prepared by coupling a partially purified Gnathostoma spinigerum-specific IgG1, MAb SK-6C4 (5 mg/ml) to CNBr-activated Sepharose 4B. Ten milliliters of approximately 0.3 mg/ml of crude soluble G. spinigerum larval antigens (GsAL3) were loaded onto the affinity column at a flow rate of about 5 ml/hour. Elution of the bound antigens was accomplished using 50 mM diethylamine-HCI containing 0.15 M NaCL, pH 11.5. The average amount of eluted antigens obtained by one passage of crude GsAL3 (1-4 mg) through 4 to 8 ml of column matrix was 143 micrograms (range, 67 - 414 micrograms). The minimal amount of purified GsAL3 detectable by ELISA using MAb SK - 6C4 (100 micrograms/ml) was 50 ng/ml. The SK - 6C4 affinity-purified GsAL3 was found to be relatively pure and immunologically specific as determined by SDS-PAGE and Western blotting, respectively.
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PMID:Purification of Gnathostoma spinigerum larval antigens by monoclonal antibody affinity chromatography. 793 40

Chronic hepatitis C virus infection can be associated with mixed cryoglobulinemia and systemic vasculitis. The pathogenesis remains poorly understood. 55 consecutive patients with chronic HCI infection (anti-HCV- and serum HCV RNA-positive) were studies prospectively. Cryoglobulinemia was detected in 28 patients (51%) with a mean cryocrit level of 2.2%. Clinical symptoms of vasculitis were encountered in six patients. Compared to those HCV-infected patients without cryoglobulinemia the following distinctive features were observed in the presence of cryoglobulinemia: increased age (p<0.02), female preponderance (p<0.002), longer-lasting HCV infection (mean of 10.7 vs. 4.7 yrs), higher prevalence of cirrhosis (42.8 vs. 0%), increased serum concentration of IgM and increased rheumatoid factor activity, decreased concentration of serum C4 (each p<0.05). The response to interferon treatment was similar in patients with and without cryoglobulinemia. When cryoprecipitates were analyzed by immunofixation, type II cryoglobulinemia was present in 1/3 and type III in 2/3 of patients. By SDS-PAGE four different proteins were demonstrable in cryoprecipitates each identified by immunoblotting as IgG and IgM heavy or light chains respectively. Cryoprecipitate IgGs were shown to react with HCV structural as well a non-structural proteins in a recombinant immunoblotting assay (RIBA). In contrast, cryoprecipitate IgMs reacted only to the HCV core protein c22-3. HCV RNA was detected in cryoprecipitates without a significant enrichment when compared to the corresponding serum or supernatant HCV RNA content. Given the monoclonality of some cryoprecipitate IgM and their reactivity to HCV core, a cross-reactivity to IgG was postulated. In fact, when performing a computer-assisted search for sequence homology, a motif within the core protein (EGLGWAGWL, conserved in HCV genotypes) was identified homologous to a sequence of IgG heavy chains. Thus, temperature-dependent affinity changes of IgM anti-HCV core (nonapeptide) and ensuing complex formation with IgG via binding to the homologous IgG sequence could be a mechanism of cryoprecipitate formation.
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PMID:Cryoglobulinemia in chronic hepatitis C virus infection: prevalence, clinical manifestations, response to interferon treatment and analysis of cryoprecipitates. 860 Jun 60

The objectives of this study were to isolate and characterize the major proteoglycans of tooth cementum in relation to the tissue's mineralization. Cementum was collected from the root apex region of bovine molars and pulverized. It was first extracted with 6M guanidine-HCI, pH 7.4 (G-extract, mineral-unassociated) and then demineralized and extracted with 0.5M EDTA (E-extract, mineral-associated). Both extracts were applied to anion exchange and then molecular sieve chromatography to isolate proteoglycans. The fractions collected were assayed for chondroitin-(CS) and keratan sulfate (KS) containing proteoglycans using the monoclonal antibodies 2-B-6 and 5-D-4, respectively. It was found that the KS was the major glycosaminoglycan and was enriched in the G-extract fraction. The major KS fraction was then applied to 7.5% SDS-PAGE. The major broad band (69 kDa) was 5-D-4 positive in Western blot analysis and separated into two bands (46 kDa and 50 kDa) after treatment with keratanase II and endo-beta-galactosidase. These two proteins were transfered to PVDF membrane and analyzed for amino acid sequence. The results showed the major band (46 kDa) to be lumican and the minor (50 kDa) fibromodulin. In addition, based on the immunohistochemical study using a number of mono- and polyclonal antibodies including 5-D-4, anti-lumican core protein as well as anti-fibromodulin core protein antibodies, the KSPGs were found to be located almost exclusively in nonmineralized portions of cementum such as precementum and the pericementocyte area. These biochemical as well as immunohistochemical data suggest that the major KSPGs of cementum, lumican and fibromodulin, have a specific tissue distribution and may have regulatory roles in cementum mineralization.
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PMID:Differential distribution of lumican and fibromodulin in tooth cementum. 890 73

The proteolytic activity of plasmin on soluble caprine beta-casein (CN) was studied in 50 mM Tris.HCI buffer, pH 8.0, at 37 degrees C. Electrophoretic studies showed that hydrolysis of this protein results in an electrophoretic pattern that is similar to the pattern obtained from plasmin hydrolysis of bovine beta-CN (gamma-CN and complementary N-terminal fragments), suggesting that plasmin probably attacks the same regions that are susceptible to cleavage in bovine beta-CN. As determined by SDS-PAGE, the gamma-like components of caprine milk consisted of two fragments with relative molecular mass of 9200 and two with relative molecular mass of 21,400 that could differ in the level of phosphorylation. Apparently, the high molecular mass components are homologous to bovine beta-CN (f 29-209) (gamma 1-CN), and the low molecular mass components are homologous to bovine beta-CN (f 106-209) and beta-CN (f 108-209) (gamma 2- and gamma 3-CN). Complementary N-terminal fragments had values for molecular masses in the range 13,600 to 8500 and urea-PAGE patterns that were more complex than those obtained in bovine casein because of the different phosphorylation levels in caprine beta-CN. These fragments were also present in the hydrolysate of whole caprine casein that had been treated with plasmin.
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PMID:Hydrolysis of caprine beta-casein by plasmin. 936 Nov 97

Several proteins in the bone matrix have been implicated in the regulation of mineral crystal formation and growth. To investigate the relationships between these proteins and the mineral phase at various stages of mineral maturation, fetal porcine calvariae and long bones were fragmented and the particles (20 microm) separated by density gradient sedimentation into fractions of increasing density (1.8 to >2.2 g/cm3). Samples from each fraction were analyzed by X-ray diffraction to obtain the average crystal size/strain and chemical composition. Other samples were sequentially extracted, first with 4.0 mol/L guanidium hydrochloride (GuHCl) (G1), then with 0.5 mol/L ethylene-diamine tetraacetic acid (EDTA) (E), and again with 4.0 mol/L Gu-HCI (G2), for analysis of proteins in different tissue compartments. Based on the mineral density distribution and crystal size, fetal porcine bone protein content was determined for tissue residue and each extract and the protein composition analyzed by sodium dodecyl-polyacrylamide gel electrophoresis (SDS-PAGE). Although the insoluble organic matrix decreased with mineral density the collagen and protein content remained fairly constant, representing approximately 10% of the tissue weight, except in the highest density fraction. Whereas the total extractable protein, representing predominantly noncollagenous proteins, did not show density-related differences, differences were observed for individual proteins on SDS-PAGE. Consistent with their presence in osteoid, the content of bone sialoprotein (BSP), tyrosine-rich acidic matrix protein (TRAMP), and a series of small proteins with cell attachment properties in the G1 extract decreased with mineral density, whereas TRAMP and BSP were increased in G2 extracts. Mineral-associated proteins, including alpha2HS-glycoprotein, BSP, osteopontin (OPN), and osteocalcin, increased with mineral density, whereas secreted protein acidic and rich in cysteine (SPARC)/osteonectin, and some minor proteins, appeared to decrease. Differences of individual proteins within and between the calvarial and long bones could be related to the role of these proteins in the formation and maturation of hydroxyapatite crystals. Collectively, these studies demonstrate mineral density-associated changes in protein composition that reflect a rapid maturation of mineral crystals in embryonic porcine bones.
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PMID:Relationships between bone protein and mineral in developing porcine long bone and calvaria. 1067 15


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