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)

Factor VIII (fVIII) is synthesized as a single chain having a domainal sequence A1-A2-B-A3-C1-C2. Analysis of the proteolyic cleavage of fVIII by thrombin by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) identifies three fragments designated fVIIIA1, fVIIIA2, and fVIIIA3-C1-C2 with fragment(s) derived from the B domain being difficult to visualize. The appearance of these fragments is associated with the development of coagulant activity, but the activity is labile without further apparent proteolysis. In this study, porcine fVIII was reacted with thrombin until peak coagulant activity was obtained and then subjected to cation-exchange (Mono S) high-pressure liquid chromatography. Coagulant activity was recovered in a single peak that contained all three fragments and was stable for weeks at 20 degrees C in 0.65 M NaCl/0.01 M His-HCl/0.005 M CaCl2 at pH 6.0. Analytical ultracentrifugation of activated fVIII was done to test whether all three fragments were associated. The apparent molecular weight of activated fVIII from equilibrium sedimentation increased from 148,000 to 161,000 as the loading concentration was increased from 0.06 to 0.16 mg/mL. This agrees well with the summed apparent molecular weights of fVIIIA1, fVIIIA2, and fVIIIA3-C1-C2 calculated from SDS-PAGE analysis (148,000) or from the amino acid sequence of human fVIII (159,000). This establishes the major species in the preparation as a fVIIIA1/A2/A3-C1-C2 heterotrimer and additionally indicates either weak self-association of the trimer and/or incomplete association of the individual subunits to form the trimer. Velocity sedimentation of activated fViii revealed a single boundry (S020,w = 7.2 S).(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Subunit structure of thrombin-activated porcine factor VIII. 249 50

Ethylenediaminetetraacetate and hydrochloric acid (EDTA) (HCl) extracts of cementum were fractionated by molecular sieving, ion exchange chromotography, and reverse phase high precision liquid chromatography (HPLC). Nine fractions were isolated, all of which contained serine phosphate, threonine phosphate, and high concentrations of aspartic acid (asp) and glutamic acid (glu). Five of the fractions obtained by repeated HPLC consisted of a single band by SDS-PAGE; the others contained at least one other minor component. All of the protein bands stained with both Rhodamine B and alcian blue, the latter consistent with analytical determinations that demonstrated that the phosphoprotein component contained a significant amount of carbohydrate, including neuraminic acid.
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PMID:Soluble glycosylated phosphoproteins of cementum. 250 8

Tetrahymena pyriformis was starved in 50 mM Tris-HCl, pH 7.5, at 28 degrees C. The number of cells did not change appreciably under the starvation conditions. Nuclear proteins of unstarved cells and cells starved for 1, 2, 4, and 7 d were analyzed by SDS-polyacrylamide gel electrophoresis. Most of the large amount of nonhistone proteins present in the unstarved cell nucleus disappeared with the starvation time. However, the relative amounts of the high mobility group protein and histones did not change appreciably. On the other hand, a protein with a molecular weight of ca. 16,000 gradually accumulated in the nucleus on starvation. This protein was extracted with 0.25 M HCl, but was not soluble in 0.5 M perchloric acid. The amino acid composition and molecular weight of this protein were similar to those of HMG protein LG-2 of T. thermophila. Some lysyl endopeptidase peptides of this protein were found to have amino acid sequences present in LG-2, thus we tentatively named it an LG-2-like protein.
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PMID:A protein that accumulates during starvation in Tetrahymena nuclei. 251 83

A new general method for the determination of neomycin phosphotransferase (NPT) II (EC 2.7.1.95) activity in cell extracts after separation in SDS-polyacrylamide gels is described. The enzymatic activity of NPT II is restored after SDS-polyacrylamide gel electrophoresis by incubating the gel for 3 h (20 mM Tris-HCl buffer, pH 7.4). The enzymatic activity is determined by in situ phosphorylation of aminoglycoside antibiotics bound to solid supports and brought into direct contact with the gel surface. A novel, mechanically stable, negatively charged matrix was synthesized for use in this solid phase enzyme assay and compared to phosphocellulose and carboxymethylcellulose paper. This new method allows the easy and exact determination of the molecular weight of any fusion protein with NPT II by assaying the position of the enzymatic activity in the gel and a consecutive immunological reaction following protein transfer onto nitrocellulose membranes.
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PMID:A new neomycin phosphotransferase II solid phase assay in combination with polyacrylamide sodium dodecylsulphate gel electrophoresis. 255 May 36

We have purified a cruciform DNA resolving endonuclease (Endo X3) greater than 1000-fold from crude extracts of mitotically growing Saccharomyces cerevisiae. The enzyme shows high specificity for DNAs with secondary structures and introduces characteristic patterns of staggered 'nicks' in the immediate vicinity of the structure. The following substrates were analyzed in detail: (i) naturally occurring four-way X junctions in cruciform DNA of a supercoiled plasmid; (ii) synthetic four-way X junctions with arms of 9 bp; (iii) synthetic three-way Y junctions with arms of 10 bp; and (iv) heteroduplex loops with 19 nucleotides in the loop. Cleavages were always found in the double stranded portion of the DNA, located immediately adjacent to the junction of the respective structure. The Endo X3 induced cleavage patterns are identical or very similar to the cleavage patterns induced in the same substrates by endonuclease VII (Endo VII) from phage T4. Furthermore, the activity of Endo X3 is completely inhibited in the presence of anti-Endo VII antiserum. Endo X3 has an apparent mol. wt of 43,000 daltons, determined by gel filtration and of approximately 18,000 daltons in SDS--polyacrylamide gels. Maximum activity of the enzyme was obtained in the presence of 10 mM MgCl2 at 31 degrees C in Tris-HCl buffer over a broad pH range with a maximum approximately 8.0. About 70% of maximal activity was obtained when Mg2+ was replaced by equimolar amounts of Mn2+ or Ca2+.
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PMID:Cruciform cutting endonucleases from Saccharomyces cerevisiae and phage T4 show conserved reactions with branched DNAs. 255 68

The present study attempts to explore the newly synthesized phosphoproteins secreted by osteoblast-like cells incubated with 1,25(OH)2D3. The phosphoproteins, which are non-collagenous proteins, may possess the ability to regulate bone mineral solubility. An osteoblast-enriched cell population isolated from 2 day-old rat calvaria by sequential enzymatic digestion was cultured in a defined medium containing dialized fetal calf serum protein (FCSP, 2 mg/ml) with 1, 5 and 10 x 10(-9)M 1,25(OH)2D3. At confluence, 32Pi (Na2H32PO4, NEX-011) was added for 24 hr. The medium proteins were precipitated by cold 10% TCA, dissolved in 15 mM Tris-HCl, pH 7.4 containing 7 M urea and chromatographed on hydroxyapatite columns (Bio-Rad, HTP). After stepwise elution with 6 mM, 0.1, 0.5 and 1.5 M Pi Hepes buffer pH 7.4 containing 3 M urea and 5 mM levamisole, the phosphoproteins were applied to 10% SDS-PAGE and autoradiographed. The 32Pi incorporated phosphoproteins of 75K, 66K, 58K, 42K, 38K, 24K, 22K, 19K, 15.5K, 13K and 3.5-10K molecular weight which were bound on a hydroxyapatite column were identified on autoradiograms of SDS-PAGE. The synthesis of 19K phosphoprotein was stable. However the synthesis of 75K, 66K, 38K and 15.5K phosphoproteins were increased by 1,25(OH)2D3. Therefore, 1,25(OH)2D3 induced phosphoproteins synthesized in rat calvarial osteoblasts, which can bind tightly on hydroxyapatite, may regulate the solubility of bone mineral and play a role in maintaining a blood/bone equilibrium.
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PMID:[Phosphoproteins biosynthesis induced by 1,25(OH)2D3 in the rat calvarial osteoblasts]. 256 75

Pennisetin, the alcohol soluble storage protein of pearl millet (Pennisetum americanum), was isolated in a homogeneous state. The intrinsic viscosity [n] of this protein was found to be in the range of 16.5-17.7 ml/g in 70% (v/v) aqueous ethanol. The [eta] changed marginally when temperature was increased from 20 to 70 degrees C and also in the presence of 10 mM NaCl. The data indicated that pennisetin was a rigid, rod shaped asymmetric hydrodynamic particle with molecular dimensions in the range of 301 x 14.4 A - 317.7 x 14.2 A. During denaturation with guanidine hydrochloride (Gdn.HCl), the intrinsic viscosity of pennisetin increased from 16 to 25ml/g with a mid point at 3.6 M of the denaturant. The native protein structure was unfolded in 6 M Gdn.HCl as shown by the exposure of aromatic amino acid residues buried in the native state and this transition was found to be reversible. The intrinsic viscosity of pennisetin in 5.9 M Gdn.HCl corresponded to Mr 25,000 which was comparable to that determined by SDS-PAGE.
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PMID:Viscometric characterization of pennisetin from pearl millet. 259 Feb 31

The major, non-amelogenin protein component (enamelin) present in EDTA or EDTA-GU HCl extracts of developing bovine enamel has a molecular weight of approximately 67 kD and an amino acid composition rich in asp, glu, ala, leuc and lys. Elution of this component from 1.5 and 3.0 mm thick strips SDS-PAGE gels and subsequent analyses show that the component selectively reacts with polyclonal antibody to bovine serum albumin. Absolute identification of this major enamelin component as serum albumin is established by an amino acid sequence of the first 40 N-Terminal amino acids which was found to be identical to bovine serum albumin. In addition to albumin, alpha-2 HS glycoprotein was also identified in the same extracts by Western blotting against a monospecific polyclonal antibody against human alpha-2 HS glycoprotein.
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PMID:Major "enamelin" protein in enamel of developing bovine teeth is albumin. 259 62

A DNA polymerase alpha-primase complex, which had been purified by means of immunoaffinity column chromatography, showed little activity in a reaction mixture composed of Tris-HCl buffer, but showed full activity in potassium phosphate buffer. It was found that potassium ion is required for the reaction by the immunoaffinity-purified enzyme. On the other hand, the DNA polymerase alpha purified by the orthodox biochemical method showed full activity in both buffer systems. A protein factor, which could restore the activity of immunoaffinity-purified DNA polymerase alpha-primase complex in the potassium-free reaction mixture, was separated from biochemically purified DNA polymerase alpha. The factor, designated as factor T, was stable to heat up to 70 degrees C, but was sensitive to trypsin. It sedimented at about 4S through a glycerol gradient. SDS-polyacrylamide gel electrophoresis revealed two polypeptide bands at 56 and 54 kDa. By immunoprecipitation, the factor T was shown to be physically associated with DNA polymerase alpha-primase complex. The stimulation was also observed with poly[d(A-T)], primed M13 DNA, and heat-denatured DNA.
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PMID:A novel stimulating protein of mammalian DNA polymerase alpha. 260 93

To study how different domains of the muscle-specific intermediate filament protein, desmin, contribute to its polymerization, two of its CNBr fragments were examined as to their oligomeric structure under assembly conditions. One of these, D88, covers residues 1-88 and represents almost the entire headpiece; the other, D109, covers residues 145-254, and includes the entire Helix 1B and part of linker L12 of the intact molecule. Chemical cross-linking followed by SDS-PAGE, and analytical gel filtration, revealed that in 10 mM Tris-HCl, pH 8.5, conditions that favor tetramerization of intact desmin D88 formed only dimers. D109, on the other hand, formed primarily a dimeric species but low levels of trimeric and tetrameric species were also detectable. These data are consistent with the proposal that, during assembly of intact protein molecules into IF, the headpiece and Helix 1 contribute to dimerization of two polypeptides into a parallel, in-register coiled-coil. However, additional interactions, including headpiece-to-rod binding and hydrophobic interaction along the entire rod domain, are required to stabilize the tetramers and full-size IF.
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PMID:Assembly properties of two CNBr fragments of avian desmin that correspond to the headpiece domain and helix 1B. 261 Jun 80


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