scholarly journals Bovine cartilage types VI and IX collagens. Characterization of their forms in vivo

1989 ◽  
Vol 262 (3) ◽  
pp. 753-761 ◽  
Author(s):  
S Ayad ◽  
A Marriott ◽  
K Morgan ◽  
M E Grant
Keyword(s):  
Molecular Mass ◽  
Proteinase Inhibitors ◽  
Guanidinium Chloride ◽  
Type Vi Collagen ◽  
Pepsin Digestion ◽  
Molecular Masses ◽  
Biosynthetic Studies ◽  
Bovine Cartilage

1. Collagens were extracted from bovine cartilage by 4 M-guanidinium chloride in the presence of proteinase inhibitors and identified by immunoblotting with specific anti-collagen sera. 2. The collagens retained their native conformations (shown by the resistance of their triple-helical domains to pepsin digestion), and the molecular masses of their component alpha-chains indicated that the chains were intact. 3. Type VI collagen was extracted as a large-molecular-mass disulphide-bonded aggregate composed of components of molecular mass 140 kDa and 200-240 kDa, and was therefore similar to type VI collagen identified in noncartilaginous tissues. Immunoblotting established the 200-240 kDa components as intact forms of the alpha 3(VI) chain. 4. Type IX collagen consisted of three clearly separable components of molecular mass 84 kDa, 72 kDa and 66 kDa, which were assigned to the alpha 1(IX)-, alpha 3(IX)- and alpha 2(IX)-chains respectively, and a large proportion of this collagen had no covalently bound glycosaminoglycan attached to the alpha 2(IX)-chain. 5. Differences between the type IX collagen extracted from bovine cartilage and that identified in biosynthetic studies on chick cartilage are discussed.

Characterization of dissolved material during the initial phase of softwood kraft pulping

TAPPI Journal ◽  
2013 ◽  
Vol 12 (1) ◽  
pp. 37-43 ◽  
Author(s):  
HANNU PAKKANEN ◽  
TEEMU PALOHEIMO ◽  
RAIMO ALÉN
Keyword(s):  
Mass Transfer ◽  
Molecular Mass ◽  
Initial Phase ◽  
Degradation Products ◽  
Kraft Pulping ◽  
Reaction Products ◽  
Cooking Temperature ◽  
Molecular Masses ◽  
Aliphatic Carboxylic Acids

The influence of various cooking parameters, such as effective alkali, cooking temperature, and cooking time on the formation of high molecular mass lignin-derived and low molecular mass carbohydrates-derived (aliphatic carboxylic acids) degradation products, mainly during the initial phase of softwood kraft pulping was studied. In addition, the mass transfer of all of these degradation products was clarified based on their concentrations in the cooking liquor inside and outside of the chips. The results indicated that the degradation of the major hemicellulose component, galactoglucomannan, typically was dependent on temperature, and the maximum degradation amount was about 60%. In addition, about 60 min at 284°F (140°C) was needed for leveling off the concentrations of the characteristic reaction products (3,4-dideoxy-pentonic and glucoisosaccharinic acids) between these cooking liquors. Compared with low molecular mass aliphatic acids, the mass transfer of soluble lignin fragments with much higher molecular masses was clearly slower.

Purification and characterization of a lectin from Xanthomonas campestris NCIM 5028

1996 ◽  
Vol 42 (6) ◽  
pp. 609-612 ◽  
Author(s):  
Bhagyashree Joshi ◽  
Jayant M. Khire ◽  
Hephzibah SivaRaman ◽  
M. Islam Khan
Keyword(s):  
Molecular Mass ◽  
Host Plant ◽  
Xanthomonas Campestris ◽  
Simple Procedure ◽  
Gel Filtration ◽  
Purification And Characterization ◽  
Ammonium Sulphate Precipitation ◽  
Molecular Masses ◽  
Lectin Purification

A lectin was isolated from culture filtrates of Xanthomonas campestris NCIM 5028, by a simple procedure of hydrophobic chromatography on phenyl-Sepharose after ammonium sulphate precipitation. The lectin was a heterodimer, with subunit molecular masses of 30 000 and 28 000. Gel filtration on S-300 column, calibrated with markers, showed its molecular mass to be approximately 70 000. Its isoelectric point was 7.2. The agglutination of the rabbit erythrocytes by the lectin was inhibited by fetuin glycopeptides and host plant (Brassica oleracea) extracts.Key words: Xanthomonas campestris, lectin, purification.

scholarly journals Localization and characterization of two structurally different forms of acetyl-CoA carboxylase in young pea leaves, of which one is sensitive to aryloxyphenoxypropionate herbicides

1994 ◽  
Vol 300 (2) ◽  
pp. 557-565 ◽  
Author(s):  
C Alban ◽  
P Baldet ◽  
R Douce
Keyword(s):  
Molecular Mass ◽  
Subunit Structure ◽  
Acetyl Coa Carboxylase ◽  
Leaf Extracts ◽  
Acetyl Coa ◽  
Native Molecular Mass ◽  
Molecular Masses ◽  
Minor Form ◽  
A Minor

Young pea leaves contain two structurally different forms of acetyl-CoA carboxylase (EC 6.4.1.2; ACCase). A minor form, which accounted for about 20% of the total ACCase activity in the whole leaf, was detected in the epidermal tissue. This enzyme was soluble and was purified to homogeneity from young pea leaf extracts. It consisted of a dimer of two identical biotinyl subunits of molecular mass 220 kDa. In this respect, this multifunctional enzyme was comparable with that described in other plants and in other eukaryotes. A predominant form was present in both the epidermal and mesophyll tissues. In mesophyll protoplasts, ACCase was detected exclusively in the soluble phase of chloroplasts. This enzyme was partially purified from pea chloroplasts and consisted of a freely dissociating complex, the activity of which may be restored by combination of its separated constituents. The partially purified enzyme was composed of several subunits of molecular masses ranging from 32 to 79 kDa, for a native molecular mass > 600 kDa. One of these subunits, of molecular mass 38 kDa, was biotinylated. This complex subunit structure was comparable with that of microorganisms and was referred to as a ‘prokaryotic’ form of ACCase. Biochemical parameters were determined for both ACCase forms. Finally, both pea leaf ACCases exhibited different sensitivities towards the grass ACCase herbicide, diclofop. This compound had no effect on the ‘prokaryotic’ form of ACCase, while the ‘eukaryotic’ form was strongly inhibited.

scholarly journals The biosynthesis of GDP-d-arabinopyranose in Crithidia fasciculata: characterization of a d-arabino-1-kinase activity and its use in the synthesis of GDP-[5-3H]d-arabinopyranose

1995 ◽  
Vol 311 (1) ◽  
pp. 307-315 ◽  
Author(s):  
P Schneider ◽  
A Nikolaev ◽  
M A Ferguson
Keyword(s):  
Molecular Mass ◽  
Kinase Activity ◽  
Substrate Inhibition ◽  
Size Exclusion ◽  
Crithidia Fasciculata ◽  
Optimum Ph ◽  
Exclusion Chromatography ◽  
Trypanosomatid Parasites

GDP-D-arabinopyranose (GDP-D-Ara) is the precursor of the uncommon D-arabinopyranose residues present in the glycoconjugates of a few trypanosomatid parasites. Biosynthetic labelling experiments with Crithidia fasciculata showed that GDP-D-Ara could be labelled with [3H]D-Ara, [2-3H]D-Glc and [6-3H]D-Glc, but not with [1-3H]D-Glc, suggesting that D-Ara can be either taken up directly by the parasite or derived from D-Glc through a pathway involving the loss of carbon C-1. In vivo pulse-chase experiments indicated that D-Ara was sequentially incorporated into D-Ara-1-PO4 and GDP-D-Ara prior to transfer to the acceptor glycoconjugate, lipoarabinogalactan. An MgATP-dependent D-arabino-1-kinase activity present in soluble extracts of C. fasciculata was purified away from phosphatase activities by size-exclusion chromatography. The D-arabino-1-kinase had an apparent molecular mass of 600 kDa, a neutral optimum pH, and displayed substrate inhibition at D-Ara concentrations above 100 microM. It had a KmATP of 1.7 mM and a KmAra of 24 microM. Competition studies indicated that the orientation of every single hydroxyl residue was important for D-Ara recognition by the enzyme, but that methyl or hydroxymethyl groups could be tolerated as equatorial substituents on C-5 of D-Ara. The partially purified D-arabino-1-kinase activity was used in the chemico-enzymic synthesis of GDP-[5-3H]D-Ara from [6-3H]D-GlcN.

scholarly journals Purification and partial characterization of the major cell-associated heparan sulphate proteoglycan of rat liver

1991 ◽  
Vol 273 (2) ◽  
pp. 415-422 ◽  
Author(s):  
M Lyon ◽  
J T Gallagher
Keyword(s):  
Molecular Mass ◽  
Proteinase Inhibitors ◽  
Gradient Centrifugation ◽  
Heparan Sulphate ◽  
Exchange Chromatography ◽  
Guanidinium Chloride ◽  
Apparent Homogeneity ◽  
Core Proteins ◽  
Sds Page ◽  
Triton X

Heparan sulphate proteoglycans were solubilized from whole rat livers by homogenization and dissociative extraction with 4 M-guanidinium chloride containing Triton X-100 and proteinase inhibitors. The extract was subjected to trichloroacetic acid precipitation and the proteoglycan remained soluble. This was then purified to apparent homogeneity by a combination of (a) DEAE-Sephacel chromatography, (b) digestion with chondroitinase ABC followed by f.p.l.c. Mono Q ion-exchange chromatography, and (c) density-gradient centrifugation in CsCl and 4 M-guanidinium chloride. Approx. 1.5 mg of proteoglycan was obtained from 30 livers with an estimated recovery of 25%. The purified proteoglycan was eluted from Sepharose CL6B as an apparently single polydisperse population with a Kav. of 0.19 and displayed a molecular mass of greater than or equal to 200 kDa (relative to protein standards) by SDS/PAGE. Its heparan sulphate chains were eluted with a Kav. of 0.44 and have an estimated molecular mass of 25 kDa. Digestion of the proteoglycan with a combination of heparinases yielded core proteins of 77, 49 and 44 kDa. Deglycosylation using trifluoromethanesulphonic acid, though slightly decreasing the sizes, gave an identical pattern of core proteins. Electrophoretic detergent blotting demonstrated that all of the core proteins were hydrophobic and are probably integral plasma membrane molecules. The peptide maps generated by V8 proteinase digestion of the two major core proteins (77 and 49 kDa) were very similar, suggesting that these two core proteins are structurally related.

scholarly journals Characterization of SENP7, a SUMO-2/3-specific isopeptidase

2009 ◽  
Vol 421 (2) ◽  
pp. 223-230 ◽  
Author(s):  
Lin Nan Shen ◽  
Marie-Claude Geoffroy ◽  
Ellis G. Jaffray ◽  
Ronald T. Hay
Keyword(s):  
Molecular Mass ◽  
Cellular Localization ◽  
Catalytic Domain ◽  
High Molecular Mass ◽  
Ran Gtpase ◽  
Specific Protease ◽  
Isopeptidase Activity ◽  
Interfering Rna

The modification of proteins by SUMO (small ubiquitin-related modifier) plays important roles in regulating the activity, stability and cellular localization of target proteins. Similar to ubiquitination, SUMO modification is a dynamic process that can be reversed by SENPs [SUMO-1/sentrin/SMT3 (suppressor of mif two 3 homologue 1)-specific peptidases]. To date, six SENPs have been discovered in humans, although knowledge of their regulation, specificity and biological functions is limited. In the present study, we report that SENP7 has a restricted substrate specificity, being unable to process SUMO precursors and displaying paralogue-specific isopeptidase activity. The C-terminal catalytic domain of SENP7 efficiently depolymerized poly-SUMO-2 chains but had undetectable activity against poly-SUMO-1 chains. SENP7 also displayed isopeptidase activity against di-SUMO-2- and SUMO-2-modified RanGAP1 (Ran GTPase-activating protein 1) but had limited activity against SUMO-1-modified RanGAP1. in vivo, full-length SENP7 was localized to the nucleoplasm and preferentially reduced the accumulation of high-molecular-mass conjugates of SUMO-2 and SUMO-3 compared with SUMO-1. Small interfering RNA-mediated ablation of SENP7 expression led to the accumulation of high-molecular-mass SUMO-2 species and to the accumulation of promyelocytic leukaemia protein in subnuclear bodies. These findings suggest that SENP7 acts as a SUMO-2/3-specific protease that is likely to regulate the metabolism of poly-SUMO-2/3 rather than SUMO-1 conjugation in vivo.

scholarly journals Evaluation of chitosan based vaginal bioadhesive gel formulations for antifungal drugs

2014 ◽  
Vol 64 (2) ◽  
pp. 139-156 ◽  
Author(s):  
Zeynep Ay Şenyiğit ◽  
Sinem Yaprak Karavana ◽  
Bayri Eraç ◽  
Özge Gürsel ◽  
Mine Hoşgör Limoncu ◽  
...  
Keyword(s):  
Molecular Mass ◽  
Antifungal Agents ◽  
Antifungal Drugs ◽  
Miconazole Nitrate ◽  
Different Types ◽  
Molecular Masses ◽  
Distribution Studies ◽  
Gel Formulations ◽  
Econazole Nitrate

Abstract The aim of the present study was to evaluate chitosan as a vaginal mucoadhesive gel base for econazole nitrate and miconazole nitrate. To this aim, different types of chitosan with different molecular masses and viscosity properties [low molecular mass chitosan (viscosity: 20,000 mPa s), medium molecular mass chitosan (viscosity: 200,000 mPa s), high molecular mass chitosan (viscosity: 800,000 mPa s)] have been used. First, rheological studies were conducted on chitosan gels. Mechanical, syringeability and mucoadhesive properties of chitosan gels were determined. Release profiles of econazole nitrate and miconazole nitrate from chitosan gels were obtained and evaluated kinetically. In addition, anticandidal activities of formulations were determined. Finally, vaginal retention of chitosan gels in rats was evaluated by in vivo distribution studies. Based on the results, it can be concluded that gels prepared with medium molecular mass chitosan might be effectively used for different antifungal agents in the treatment of vaginal candidiosis, since it has high mucoadhesiveness, suitable mechanical and release properties with good vaginal retention

Characterization of a recombinant porcine follistatin in a heat shock expression system

2002 ◽  
Vol 82 (3) ◽  
pp. 295-304
Author(s):  
C. R. Christensen ◽  
J. Kowalski ◽  
P. J. Chedrese ◽  
V. Misra ◽  
B. Laarveld ◽  
...  
Keyword(s):  
Heat Shock ◽  
Ovarian Function ◽  
Expression System ◽  
Porcine Granulosa Cells ◽  
Molecular Masses ◽  
Estradiol 17Β

The role of follistatin in ovarian function has yet to be fully elucidated; it is present in low concentration in vivo and it is difficult to obtain suitable quantities of follistatin for characterization. We have cloned porcine follistatin cDNA in an expression system that uses the heat shock protein promoter BoHSP70. Recombinant follistatin with apparent molecular masses of 39, 46, 48, 50 kDa was expressed and secreted into culture medium at concentrations of 400–500 μg × 20 mL-1 × 150 cm-2 flask (4 × 107 cells). In ligand blots, the recombinant follistatin was shown to be immunologically similar to native follistatin and to bind to recombinant activin A. Porcine granulosa cells dissected from 1–3 mm follicles from immature gilts were cultured with recombinant follistatin or with a follistatin monoclonal antibody to examine the activity of the recombinant follistatin. At a physiologically relevant concentration of 1 μg mL-1 the recombinant porcine follistatin suppressed the accumulation of estradiol-17β (P < 0.05). There was a trend for estradiol-17β accumulation in the presence of 10 μg mL-1 of monoclonal anti-follistatin antibody (P = 0.1). This expression system consistently produced large quantities of recombinant porcine follistatin, which is immunologically and biologically similar to follistatin, and is capable of independently inhibiting estratiol-17βproduction in vitro. Key words: Porcine, follistatin, heat shock promoter, glycoprotein, ovary, estradiol

scholarly journals Isolation and characterization of human cervical-mucus glycoproteins

1983 ◽  
Vol 211 (1) ◽  
pp. 13-22 ◽  
Author(s):  
I Carlstedt ◽  
H Lindgren ◽  
J K Sheehan ◽  
U Ulmsten ◽  
L Wingerup
Keyword(s):  
Density Gradient ◽  
Proteinase Inhibitors ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Disc Electrophoresis ◽  
Guanidinium Chloride ◽  
Molecular Weights ◽  
Isolation And Characterization ◽  
Mucus Glycoproteins

Mucus glycoproteins (mucins) were extracted from human cervical pregnancy mucus by 6 M-guanidinium chloride in the presence of proteinase inhibitors. Purification was subsequently achieved by isopycnic density-gradient centrifugation in CsCl/ guanidinium chloride gradients. The purified macromolecules represented approx. 85% of the total and were devoid of nucleic acids and proteins, as judged by analytical density-gradient centrifugation, disc electrophoresis and u.v. spectroscopy. Sedimentation-velocity centrifugation revealed a single unimodal peak with S20,W 50.1S in 0.2M-NaCl and 37.0S in 6 M-guanidinium chloride. Molecular weights obtained by light-scattering were 9.7 × 10(6) and 5.9 × 10(6) in 0.2M-NaCl and 6 M-guanidinium chloride respectively. The chemical analyses were typical of those of epithelial mucins. The macromolecules contained approx. 20% (w/w) of protein, and 65% (w/w) was accounted for as carbohydrate. Serine and threonine constituted 32 mol/100 mol and proline 10 mol/100 mol of the amino acids. The major sugars found were N-acetylglucosamine (12.8%), N-acetylgalactosamine (9.7%), galactose (18.7%), sialic acid (15.0%) and fucose (7.5%).

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