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)

Both 7 alpha- and 7 beta-hydroxysteroid dehydrogenases (HSDH) were induced by either chenodeoxy-(CDC) or deoxycholic (DC) acid in C. absonum. 7 beta-HSDH was partially purified 35-fold from CDC-induced cultures of C. absonum by Procion Red (PR) affinity chromatography and high performance liquid chromatography (HPLC) using a TSK 3000 SW gel filtration column. A relative molecular weight of 200 K was estimated for 7 beta-HSDH using Sephacryl S-300 chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the 35-fold purified 7 beta-HSDH showed six polypeptides in the molecular weight range of 40-50 K. Induction of cultures of C. absonum with CDC or DC (0.4 mM) also resulted in the differential synthesis of at least five new polypeptides with molecular weights of 94 K, 42 K, 32 K, 21 K, and 16 K. The 16 K polypeptide was induced by DC but not by CDC. SDS-PAGE of Triton X-100-solubilized membranes from these extracts revealed the presence of a new membrane-associated polypeptide of molecular weight 80 K. The soluble inducible polypeptides were eliminated during purification of the 7 alpha- and 7 beta-HSDH and, therefore, are not required for these enzyme activities. It is proposed that this organism synthesized 7 alpha- and 7 beta-HSDH as well as a series of other proteins in response to bile acids which may, in the absence of the dehydrogenases, be toxic to C. absonum. The HSDH's catalyze the epimerization of chenodeoxycholic acid to ursodeoxycholic acid, which is less toxic than the chenodeoxycholic acid. The other proteins may assist the survival of the organism in a high bile acid environment by mechanisms not yet understood.
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PMID:Separation of 7 alpha- and 7 beta-hydroxysteroid dehydrogenase activities from clostridium absonum ATCC# 27555 and cellular response of this organism to bile acid inducers. 657 44

A simple and rapid method has been developed for the separation of apolipoproteins in high density lipoprotein (HDL) fractions by high performance liquid chromatography (HPLC) with gel permeation columns (G3000SW TSK GEL). The HPLC pattern monitored by A280 for a mixed solution of the HDL fraction (10 microliter) and an eluent buffer (200 microliter, 0.1 M sodium phosphate buffer containing 0.1% SDS) incubated at 60 degrees C for 5 min showed two completely separated peaks which corresponded to the major components of human HDL, apolipoprotein A-I and A-II. Moreover, quantitation of apolipoprotein A-I by our method was found to correlate well with that by a single radial immunodiffusion (SRID) assay.
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PMID:Simple method for analysis of apolipoproteins in high density lipoproteins by high performance liquid chromatography. 663 Jan 80

Rat liver cytosol contains proteins which in the presence of low-molecular-weight metabolites modulate activities of membrane-bound enzymes of cholesterol biosynthesis. In the preceding paper, we identified Z-protein as a mediator in fatty acyl-CoA modulation of microsomal cholesterol synthetic and metabolizing enzymes. In this communication, we describe a second cytosolic protein which displays cholesterol-exchange activity. Purification of the protein to over 10000-fold and homogeneity has been achieved by gel permeation HPLC on an analytical Spherogel TSK-2000 SW column. Elution of both a single peak of active protein and one SDS-polyacrylamide gel electrophoresis species upon HPLC-purification suggests that homogeneous protein aggregates, with loss of exchange activity. In addition to stimulating microsomal enzymes of sterol synthesis, incubations of microsomes with cholesterol-containing liposomes and the protein consistently yields a 2-3-fold stimulation of microsomal acyl CoA: cholesterol acyltransferase activity. Under similar incubation conditions the protein enhances only slightly the extent of inhibition of microsomal hydroxymethylglutaryl-CoA reductase by liposomal cholesterol. The protein also catalyzes net transfer of cholesterol between membranes of different cholesterol content. The lipid-transfer protein and another cytosolic protein, also implicated in the regulation of sterol synthetic enzymes, appear identical. Regulation of activities of several membrane-bound enzymes of cholesterol metabolism in which the lipid-transfer protein and cytosolic Z-protein modulate uptake of lower-molecular-weight water-insoluble and water-soluble effectors, respectively, is discussed.
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PMID:Cytosolic modulators of activities of microsomal enzymes of cholesterol biosynthesis. Purification and characterization of a non-specific lipid-transfer protein. 683 Aug 31

Agglutinins from Tachypleus (Tachypleus tridentatus, the Japanese horseshoe crab) hemolymph were isolated by affinity chromatography on BSM-coupled Sepharose 4B. The agglutinins showed multiple species and were composed of eight heterogeneous subunits with molecular weights of 45,000, 42,000, 41,000, 39,000, 33,000, 29,000, 27,000, and 22,000 as determined by SDS-polyacrylamide gel electrophoresis. The affinity-isolated agglutinins were fractionated into four groups by gel filtration on a Fractogel TSK (Toyopearl) HW-65 column, and these were designated as Tachypleus tridentatus agglutinin (TTA)-I, -II, -III, and -IV in the order of elution. These agglutinins were demonstrated to be heterogeneous as judged by their specificity towards horse erythrocytes, subunit structures, and immunological properties. TTA-III showed a potent agglutination activity towards horse erythrocytes and was further purified by gel filtration on a Cellulofine GC-700 column. The purified TTA-III is a highly purified (46,000-fold) protein composed of homogeneous subunits (Mr, 42,000) as judged by SDS-polyacrylamide gel electrophoresis and immunological analysis.
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PMID:Agglutinins in the horseshoe crab hemolymph: purification of a potent agglutinin of horse erythrocytes from the hemolymph of Tachypleus tridentatus, the Japanese horseshoe crab. 688 38

Two enzymes with phospholipase D activity were purified from Streptomyces strains (PMF and PM43) by column chromatography on Fractogel TSK CM-650(S), Sephadex G-100 and Fractogel EMD DEAE-650(M). The purified preparations were found to be homogeneous by SDS-PAGE, capillary electrophoresis and analytical gel filtration. The molecular masses, assessed by MALDI-MS spectrometry, were 53.864 kDa for PMF and 54.147 kDa for PM43. The isoelectric point was 9.1 for both enzymes. The enzymes were most active at around 60 degrees C and stable between pH 4 and 9 and below 50 degrees C. The pH optima were between 4 and 6 for PMF and between 6 and 7 for PM43. Both phospholipases displayed high transphosphatidylation activity but PMF was more selective than PM43.
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PMID:Purification and properties of two phospholipases D from Streptomyces sp. 773 43

Ceramidase (CDase) catalyzes the hydrolysis of ceramides to yield sphingosine and fatty acid. In this paper, two forms of membrane-bound alkaline ceramidase, have been, for the first time, purified from guinea pig epidermis by chromatography on DEAE-cellulose, phenyl-Superose, HCA-hyroxyapatite, isoelectric focusing, Mono Q, and TSK-3000SW column. One species (CDase-I) migrated upon SDS-polyacrylamide gel electrophoresis as a single band with an apparent molecular mass of 60 kDa; the other (CDase-II) was only partially purified with apparent M(r) of about 148,000 estimated by gel filtration. The specific activities of the two species increased by 1.130- (for CDase-I) and 400-fold (for CDase-II) over the original tissue extract. The activity of both enzymes for ceramide species decreased in the order of linoleoyl > oleoyl > palmitoylsphingosine. The optimal pH for enzyme activity was approximately 7.0-9.0 for CDase-I and 7.5-8.5 for CDase-II. Interestingly, both enzymes were inhibited by the reaction product sphingosine with a concentration for half-maximal inhibition (ID50) of 100-130 microM, compared to the apparent kinetic parameters with CDase-I (Km = 90 microM, Vmax = 0.62 unit) and CDase-II (Km = 140 microM, Vmax = 0.50 units). Some lipids, such as phosphatidylcholine and sphingomyelin, are also inhibitory with IC50 values of 50-250 microM, suggesting well controlled CDase activity by sphingolipid metabolites. These studies begin to elucidate a regulatory mechanism for the balance of the ratio of ceramide/sphingosine which can serve as an intracellular effector molecule in epidermis.
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PMID:Purification and biochemical characterization of membrane-bound epidermal ceramidases from guinea pig skin. 775 19

alpha-L-Arabinofuranosidase was purified from a cell-free extract of Aspergillus niger 5-16 by chromatographies on DEAE-Toyopearl, SP-Toyopearl, Ultro-gel AcA 44, Mono P, and TSK-Gel G3000SW. The final preparation thus obtained showed a single band on SDS-polyacrylamide gel electrophoresis. The molecular weight and isoelectric point were 67,000 by SDS-polyacrylamide gel electrophoresis and pH 3.5 by isoelectric focusing. The alpha-L-arabinofuranosidase contained amino acids in the order of Asx > Gly > Ala > Thr > Glx = Ser. The enzyme had maximum activity at pH 4.0 and 60 degrees C, and was stable from pH 4 to 7 and at temperatures up to 30 degrees C. The enzyme activity was not affected considerably by either metal ions or chemical reagents. The enzyme released arabinose from p-nitrophenyl-alpha-L-arabinofuranoside, O-alpha-L-arabinofuranosyl-(1-->3)-O-beta-D-xylopyranosyl-(1-->4)-D- xylopyranose, and arabinan, but not from O-beta-D-xylopyranosyl-(1-->4)-O-[alpha-L-arabinofuranosyl- (1-->3)]-O-beta-D-xylopyranosyl-(1-->4)-D-xylopyranose, O-beta-D-xylopyranosyl-(1-->2)-O-alpha-L-arabinofuranosyl-(1-->3)-O-beta -D- xylopyranosyl-(1-->4)-O-beta-D-xylopyranosyl-(1-->4)-D-xylopyranose, gum arabic, or arabinoxylan. The limit of hydrolysis of arabinan was about 58% even when the enzyme was sufficiently in excess.
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PMID:Purification and some properties of intracellular alpha-L-arabinofuranosidase from Aspergillus niger 5-16. 776 88

One strain (MO-5) of fungi producing alpha-D-xylosidase was isolated from sake koji by a method using glucose oxidase. MO-5 was identified as an Aspergillus flavus strain, produced no aflatoxin (B1, B2, G1, and G2). Two different types of alpha-D-xylosidases were detected in the cell-free extract. One of the enzymes (alpha-D-xylosidase I) was purified to an electrophoretically pure state by successive chromatography on Q-Sepharose, Phenyl Superose, PL-SAX, and TSK-gel G3000SWXL. The purified enzyme hydrolyzed p-nitrophenyl alpha-D-xylopyranoside (alpha-p-NPX), methyl alpha-D-xylopyranoside, isoprimeverose [alpha-D-xylopyranosyl-(1-->6)-D- glucopyranose], and xyloglucan oligosaccharide. The activity of this enzyme was highly specific for alpha-D-xylosidic linkages. The apparent Km and Vmax of the enzyme for alpha-p-NPX and isoprimeverose were 1.32 mM and 4.4 mumol/min/mg protein, and 4.00 mM and 58.8 mumol/min/mg protein, respectively. This enzyme had an apparent molecular weight of 100,000 by SDS-polyacrylamide gel electrophoresis and 400,000 by gel filtration chromatography (TSK-gel G3000SWXL). The enzyme showed the highest activity at pH 4.5 and 45 degrees C, and was stable from pH 4.5 to 6.0 and at temperatures up to 45 degrees C. The activity was inhibited by SDS and Hg2+, and slightly by Cu2+ and Fe3+. This enzyme showed transfer action at high concentrations of isoprimeverose and transfer products were detected, and it had transxylosylation activity on maltose from isoprimeverose as a donor, too.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Isolation of Aspergillus flavus MO-5 producing two types of intracellular alpha-D-xylosidases: purification and characterization of alpha-D-xylosidase I. 776 13

Alpha-D-Xylosidase II activity from Aspergillus flavus MO-5 was increased roughly 5- to 10-fold by use of xylose instead of methyl alpha-D-xylopyranoside (alpha-MX) as a carbon source. The enzyme was purified to an electrophoretically pure state by successive chromatography on Q-Sepharose, Phenyl Superose, PL-SAX, and TSK-gel G3000SWXL. The purified enzyme hydrolyzed isoprimeverose [alpha-D-xylopyranosyl-(1-->6)-D-glucopyranose] and p-nitrophenyl alpha-D-xylopyranoside (alpha-p-NPX), but not alpha-MX or xyloglucan oligosaccharide. The apparent Km and Vmax of the enzyme for alpha-p-NPX and isoprimeverose were 0.97 mM and 28.0 mumol/min/mg protein, and 47.62 mM and 2.0 mumol/min/mg protein, respectively. This enzyme had an apparent molecular weight of 67,000 by SDS-polyacrylamide gel electrophoresis and 180,000 by gel filtration chromatography (TSK-gel G3000SWXL). The enzyme showed the highest activity at pH 6.0 and 40 degrees C, and was stable in the pH range from 6.0 to 7.0 and at the temperatures up to 40 degrees C. The activity was inhibited by Cu2+, Zn2+, Hg2+, p-CMB, SDS, Fe3+, and N-ethylmaleimide. This enzyme had nothing in common with alpha-D-xylosidase I and four alpha-D-xylosidases reported already.
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PMID:Purification and characterization of an intracellular alpha-D-xylosidase II from Aspergillus flavus MO-5. 776 14

Ginger proteases in ginger rhizome (Zingiber officinale roscoe) were extracted from the ginger acetone powder and purified on DEAE-Sepharose and Sephadex G-75 columns. Before the purification, excess p-chloromercuribenzoate was added to the enzymes to prevent their autodigestion. The mercuribenzoate-proteases were further purified and fractionated by isoelectric focusing in Ampholine of pH 3-10 or pH 4-6. The proteases were fractionated into three components by the isoelectric focusing, having pI value of 4.5, 4.6 and 4.8 respectively. All these proteases had a molecular mass of 29,000 as measured by SDS-polyacrylamide gel electrophoresis and by TSK G2000SW XL gel chromatography. The Ampholine in the purified enzymes can quickly be removed by the gel chromatography of TSK G2000SW. Some divalent metal ions, such as Hg2+, Cu2+, Cd2+, and Zn2+, strongly inhibited these purified enzymes.
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PMID:Purification of ginger proteases by DEAE-Sepharose and isoelectric focusing. 787 61


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