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)

An alpha-amylase inhibitor (PHA-I) of the white kidney bean (Phaseolus vulgaris) was found to be composed of two kinds of subunits and they were isolated on a size-exclusion column by HPLC under denaturing conditions. The alpha-subunit was free from tryptophan and cysteine and the beta-subunit contained no methionine or cysteine. There was no marked resemblance in tryptic peptide map between these subunit polypeptides. The alpha-subunit contained 28% by weight of carbohydrate, mainly made up of high mannose-type oligosacharides, whereas the sugar moiety of the beta-subunit amounted to 7% by weight and seemed to be predominantly composed of xylomannose-type oligosaccharides. By SDS-PAGE following deglycosylation, the molecular weights of the polypeptides of alpha- and beta-subunits were shown to be 7,800 and 14,000, respectively. These values were consistent with molecular sizes obtained for alpha- and beta-subunits by gel permeation HPLC in 6 M guanidine hydrochloride. The molecular weight of the native PHA-I, 28,800, obtained by gel permeation HPLC under non-denaturing conditions, suggested a heterodimeric structure for PHA-I.
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PMID:Isolation and characterization of the subunits of Phaseolus vulgaris alpha-amylase inhibitor. 178 11

Antigen 5.1 was isolated from the most acidic fraction of castor bean allergens (CB-1A) by gel filtration on Sephadex G-75 followed by polyacrylamide gel electrophoresis (PAGE) (yield: 6.2 mg antigen 5.1/g CB-1A). This antigen was homogeneous by the criteria of PAGE, isoelectric focusing, SDS-PAGE, immunoelectrophoresis and gel filtration. The antigen has an apparent molecular mass of 12 +/- 2 kDa and an isoelectric point of pH 5.1. Antigen 5.1 lacks proline, phenylalanine, threonine and tryptophan. It was immunochemically identical to one of the three immunoprecipitation lines presented by CB-1A by the Ouchterlony technique, and was positive when tested (250 micrograms) by IgE-mediated passive cutaneous anaphylaxis in LOU.M rats.
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PMID:Isolation and partial characterization of antigen 5.1 from the seeds of Ricinus communis, L. (Var. Amarelo de Irece). 182 25

Exposure of purified human plasma fibronectin to the myeloperoxidase-H2O2-Cl- system of neutrophils or to reagent HOCl resulted in extensive changes to its primary and tertiary structures. When 1.14 microM fibronectin was exposed to 50-400 microM HOCl or 50-400 microM H2O2 plus myeloperoxidase and Cl-, there was progressive loss of tryptophan fluorescence and cysteines, and an increase in bityrosine fluorescence and carbonyl content. Analysis by SDS-PAGE indicated extensive crosslinking of the fibronectin, the crosslinks being stable under reducing conditions. The coincident increase of bityrosine fluorescence suggests that crosslinking may be largely due to intermolecular bityrosines rather than disulfides. All changes observed with the myeloperoxidase system were inhibited by azide or methionine, and were dependent upon the presence of chloride, indicating that they are mediated by HOCl. The reaction between HOCl and fibronectin resulted in the formation of long-lived chloramines. Exposure to increasing amounts of oxidant resulted in an increase in the susceptibility of fibronectin to proteolytic attack by purified neutrophil elastase. Analysis by SDS-PAGE showed a different fragmentation pattern for oxidant-treated fibronectin compared with the native protein. This suggests that regions of the molecule which were previously resistant to proteolysis were denatured to create susceptible sites for elastase. This demonstration that fibronectin is extensively modified by the myeloperoxidase system has implications for the mechanism of tissue injury by neutrophils in inflammation, since a loss of functional fibronectin would result in cell detachment and a distortion of normal tissue organization.
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PMID:Oxidative damage to fibronectin. I. The effects of the neutrophil myeloperoxidase system and HOCl. 184 32

To investigate the possible role of glycation in the onset of diabetic cataract we used calf lens crystallins as a model. After incubation with reducing sugars, the proteins were investigated by high-pressure gel permeation chromatography, SDS-PAGE and analytical ultracentrifugation. Glucose-6-phosphate incubation resulted in an increase in mean molecular weight of all crystallin fractions and the occurrence of high-molecular weight material, partly formed by disulphide bonds. The glycated crystallins showed a decrease of tryptophan fluorescence and an increase of a specific non-tryptophan fluorescence. This fluorescence was, however, not exclusively associated with the high molecular weight protein, but was present in all protein fractions.
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PMID:Glycation-induced crosslinking of calf lens crystallins. 187 6

We isolated and sequenced a cDNA clone corresponding to the entire coding sequence of rat liver lysosomal cathepsin D. The deduced amino acid sequence revealed that cathepsin D consists of 407 amino acid residues (Mr 44,608) and the 20 NH2-terminal residues seem to constitute a cleavable signal peptide after which 44 amino acid residues follow as a propeptide. Two putative N-linked glycosylation sites and aspartic acid in the active site are as well conserved as those of human lysosomal cathepsin D. In the NH2-terminal sequence analysis of two isolated heavy chains of the mature enzyme, the termini were assigned as tryptophan (118th residue) and glycine (165th or 166th residue), respectively, hence demonstrates that the two heavy chains derive from a split of the single chain of cathepsin D at position between 117th and 118th or between 164th and 165th or 165th and 166th amino acids. We conclude that cathepsin D in rat liver lysosomes is a mixture of three forms composed of a single and two two-chain forms. However, the amounts of the two two-chain forms are low compared with that of the single chain form. Densidometric determination after SDS-PAGE revealed that the two two-chain forms account for less than 5% of the single chain form. There is a 82% similarity in amino acid level between rat and human liver lysosomal cathepsin D.
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PMID:Isolation and sequencing of a cDNA clone encoding rat liver lysosomal cathepsin D and the structure of three forms of mature enzymes. 188 50

The acid release of endogenous peptides from immunoaffinity-pure human major histocompatibility complex (MHC) class II proteins HLA-DR1 is accompanied by an 18% decrease in intrinsic tryptophan fluorescence. The effect is totally reversible upon readdition of an autologous endogenous peptide fraction. High-performance size-exclusion chromatographic (HPSEC) binding and release studies with a nonfluorescent HLA-DR1-restricted influenza matrix peptide IM(18-29) prove the fact that Trp residues of the HLA protein change their fluorescence intensities. Since the far-UV circular dichroism spectra of HLA molecules before and after peptide release, DR1[NAT] and DR1[REL], show very small differences, we can rule out the breakdown of secondary structural elements under release conditions, although DR[REL] consists of disassembled alpha- and beta-subunits, as evidenced by HPSEC. Quenching of DR1[NAT] and DR1[REL] using the neutral quencher acrylamide results in a 20% increase in total accessibility of the nine-residue Trp population whereas quenching by iodide yields only a 5% increase. Both results taken together tell us that two Trp residues, preferentially ones located in apolar pockets, become accessible upon the release of peptides. The significantly smaller fluorescence enhancement upon binding IM(18-29) of DR3[REL], exclusively lacking Trp-9(beta 1), and the missing tendency to reassemble under the influence of IM(18-29) compared to DR1[REL] suggest an important role for position 9(beta 1). The region around Trp-43(alpha 1) should be responsible for the binding of IM(18-29) to the alpha-subunits of DR1 and DR3, respectively, as verified by fluorometric HPSEC and SDS-PAGE. Obviously, our findings are in total agreement with the hypothetical MHC class II model, whereafter Trp-9(beta 1) and Trp-43(alpha 1) besides Trp-61(beta 1) are constituents of the binding groove of DR1. Extending the homology to MHC class I products, we postulate the existence of three hydrophobic pockets in the binding site of DR1 with the cited Trp residues being juxtaposed to contacting apolar peptide side chains in HLA-peptide complexes. According to the deduced two-residue-contact model the minimal consensus motif for DR1-restricted peptide antigens consists of two hydrophobic residues lying 14-16 A apart in the bound state of the peptide.
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PMID:Self and foreign peptides interact with intact and disassembled MHC class II antigen HLA-DR via tryptophan pockets. 189 27

Alpha-1-proteinase inhibitor (alpha-1-PI) was isolated from goat plasma by salt fractionation, and chromatography on a DEAE-cellulose column. The inhibitor was found to be homogeneous by gel chromatography, SDS-PAGE and PAGE.Mr values by gel filtration (57 kDa), and by SDS-PAGE (52 kDa), under reducing conditions were nearly the same suggesting that the inhibitor consists of a single polypeptide chain. It contained 13.8% neutral hexose but no sialic acid residue. The values of isoionic pH, and extinction coefficient at 278 nm were 4.84, and 4.6, respectively. Fluorescence spectral properties showed tryptophan residues in the inhibitor. Solvent perturbation difference spectra suggested 74% exposure of the tryptophan residues in the native molecule. Gel filtration behaviour of the inhibitor was consistent with a Stokes radius of 3.16 nm, diffusion coefficient of 7.02 X 10(-7) cm2-sec-1 and a frictional ratio of 1.24 suggesting asymmetry and/or excessive hydration of the inhibitor molecule. Goat alpha-1-PI, unlike human alpha-1-PI was found to be potent inhibitor of bovine trypsin but a poor inhibitor of porcine pancreatic elastase. It was virtually devoid of antichymotryptic activity.
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PMID:Isolation and characterization of alpha-1-proteinase inhibitor from goat plasma. 193 Feb 50

Translation of tobacco vein mottling virus (TVMV) RNA in a wheat germ system resulted in two products that were not observed in a rabbit reticulocyte system. One of these was the N-terminal protein, based on its being the most abundant product and its migration on SDS-PAGE at about 34 kDa. The second product was similar or identical to helper component (HC) isolated from TVMV-infected plants, based on co-migration with HC on SDS-PAGE and immunoprecipitation with anti-HC antibodies. The N-terminus of this product was determined by radiochemical Edman degradation to be Ser-257 of the polyprotein. This assignment was supported by peptide mapping with a tryptophan-specific reagent. A similar cleavage was observed when tobacco etch virus was translated in a wheat germ system. Comparison with homologous regions in five other potyviruses indicated conservation of amino acid residues on both sides of the proposed cleavage site. Conversion of Phe-256 to Met, Pro, Arg, His, or Trp by site-directed mutagenesis of a TVMV RNA transcription template inhibited cleavage in the wheat germ system. These results suggest that in vitro cleavage occurs between Phe-256 and Ser-257 and that this cleavage is the same as the in vivo cleavage which liberates the N-terminus of HC.
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PMID:In vitro cleavage at or near the N-terminus of the helper component protein in the tobacco vein mottling virus polyprotein. 196 46

The present investigation evaluated amino acid utilization by 120 strains of Fusobacterium nucleatum in a chemically defined medium and attempted to relate the patterns to 3 proposed subspecies of F. nucleatum. Strains were inoculated into a chemically defined medium, with and without 2 g/l glucose, consisting of 14 inorganic salts, 21 amino acids, 23 vitamins and cofactors, and 7 purines and pyrimidines. After 7 days of anaerobic incubation, the spent culture medium, as well as the uninoculated control medium, were analyzed for amino acid content by ion chromatography. Amino acid utilization was determined by the differences in concentrations of amino acids found in inoculated and uninoculated samples. If greater than 34% of the amino acid was removed from the medium, the amino acid was considered to be utilized. Of the 21 amino acids present in the chemically defined medium, 8 amino acids, lysine, glutamine, asparagine, histidine, threonine, serine, glutamate and cysteine were consistently utilized. Four amino acids, tyrosine, tryptophan, methionine and aspartate were utilized by some strains but not others. Nine amino acids, alanine, leucine, isoleucine, glycine, valine, phenylalanine, proline, ornithine, and arginine were not utilized by any of the strains. The utilization patterns did not relate to subspecies formed on the basis of SDS-PAGE and DNA hybridization.
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PMID:Amino acid utilization by Fusobacterium nucleatum grown in a chemically defined medium. 208 74

Four fragments of ovomucoid representing its individual domains and their different combinations were prepared by peptic and cyanogen bromide cleavages of the protein. The fragments corresponding to domains I + II, II + III, I and III of the parent ovomucoid molecule, were found to be homogeneous by gel filtration and polyacrylamide gel electrophoresis in presence and absence of SDS. Various physico-chemical properties of these proteins, such as molecular weight, NH2- and COOH-terminal amino acid residues, sugar content, isoionic pH, specific extinction coefficient, fluorescence emission spectra, intrinsic viscosity, frictional coefficient, Stokes radius, diffusion coefficient and geometrical mean radius were determined. Analysis of the results on trypsin inhibitory activity of ovomucoid and its different fragments suggested that only domain II is involved in the antitryptic activity of the inhibitor. Optical characteristics of these fragments indicate that they are devoid of tryptophan residues. The hydrodynamic properties suggest that intact ovomucoid and two of its fragments (domain I + II and domain II + III) are significantly different from those of typical globular proteins and are asymmetric in nature. However, the shape of the two remaining fragments representing domains I and III of the intact protein appeared to be compact and globular. Furthermore, domain II of ovomucoid has been suggested to primarily contribute towards the apparent asymmetry in the intact protein.
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PMID:Ovomucoid domains: preparation and physico-chemical characterization. 209 Jan 11


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