Multiple forms of ribonuclease II in mouse liver: relative sizes, subcellular distributions, and pH–activity profiles

1987 ◽  
Vol 65 (1) ◽  
pp. 27-34 ◽  
Author(s):  
Scot R. Kimball ◽  
William L. Meyer
Keyword(s):  
Molecular Weight ◽  
Mouse Liver ◽  
Heavy Particle ◽  
Gel Permeation Chromatography ◽  
Apparent Molecular Weight ◽  
Total Activity ◽  
Exchange Chromatography ◽  
The Other ◽  
Multiple Forms ◽  
Small Series

Multiple forms of ribonuclease II (EC 3.1.27.5) have been resolved from extracts of crude fractions of mouse liver by ion-exchange chromatography on phosphocellulose and gel permeation chromatography. The forms are designated 6S, 6L, 5S, 5L, 4S, 4L, 3S, 3L, 2, and 1 in increasing order of apparent cationic character. The forms fall into two series of apparent molecular weight. The small series increases from molecular weight equal to 9000 for form 1 to 14 000 for form 6S. The large series increases from molecular weight equal to 22 000 for form 2 to 44 000 for form 6L. All forms have pH–activity profiles with maxima near pH 7. Activity falls to no less than 30% of this maximum at pHs 5 and 8.5. Relative to the other forms, form 1 has a higher ratio of activity in the alkaline compared with acid pH range. Form 1 is found in the cytosolic, "light" particle, and "heavy" particle fractions. The other forms are largely restricted to the heavy particle fraction. In this fraction the proportion of total activity attributable to each form generally decreases in order from form 1 down to form 6. The results are accommodated by models in which one or more gene products give rise to multiple forms of ribonuclease II by processes involving dimerization and glycosylation.

scholarly journals The multiple forms of α-D-mannosidase in human plasma

1979 ◽  
Vol 179 (3) ◽  
pp. 583-592 ◽  
Author(s):  
S Hirani ◽  
B Winchester
Keyword(s):  
Molecular Weight ◽  
Human Plasma ◽  
Concanavalin A ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Exchange Chromatography ◽  
The Other ◽  
Multiple Forms ◽  
Simple Correlation ◽  
Multiple Components

The acidic alpha-D-mannosidase in human plasma closely resembles liver acidic alpha-D-mannosidase in its affinity for concanavalin A-Sepharose, molecular weight and resolution into multiple components on DEAE-cellulose. A combination of chromatography on concanavalin A-Sepharose and gel filtration on Sephadex G-200 and Sepharose 6B suggests that four forms of intermediate alpha-D-mannosidase, which differ either in their molecular weight of affinity for concanavalin A, exist in human plasma. A practical classification and nomenclature for the multiple forms of intermediate alpha-D-mannosidase in plasma based on molecular weight and affinity for concanavalin A is proposed. Multiple forms of intermediate alpha-D-mannosidase were also observed by ion-exchange chromatography on DEAE-cellulose, but there was not a simple correlation between these forms and those obtained with the other separation procedures. The form of intermediate alpha-D-mannosidase least abundant in plasma, approx. 7% of the activity, has very similar properties to the neutral alpha-D-mannosidase in human liver. In contrast, the other three forms of intermediate alpha-D-mannosidase, which account for over 90% of the activity, do not appear to be present in liver, except perhaps in trace amounts.

Detection of inducible nitric oxide synthase inSchistosoma japonicumandS. mansoni

2004 ◽  
Vol 78 (1) ◽  
pp. 47-50 ◽  
Author(s):  
X.-C. Long ◽  
M. Bahgat ◽  
K. Chlichlia ◽  
A. Ruppel ◽  
Y.-L. Li
Keyword(s):  
Nitric Oxide ◽  
Molecular Weight ◽  
Inducible Nitric Oxide Synthase ◽  
Mouse Liver ◽  
Apparent Molecular Weight ◽  
Western Blots ◽  
Oxide Synthase ◽  
Host Tissues ◽  
Inducible Nitric Oxide

AbstractSchistosoma japonicumandS. mansoniwere tested for reactivity with an anti-inducible nitric oxide (iNOS) antibody and the distribution of iNOS was studied by immunofluorescent tests in different stages of the parasites. Reactivity was associated with the tegument in both larval schistosomes (sporocysts and cercariae) and eggs. With adult worms, the majority of the immunofluorescence was predominantly subtegumental inS. japonicumand parenchymal inS. mansoni. Fluorescence was also observed in host tissues (snails and mouse liver). In Western blots, the enzyme ofS. japonicumhad an apparent molecular weight of about 210 kDa. The possible role of worm and host iNOS in the parasite–host interrelation remains to be clarified.

scholarly journals Proteinaceous Protease Inhibitor from Lawsonia Inermis: Purification, Characterization and Antibacterial Activity

2013 ◽  
Vol 8 (10) ◽  
pp. 1934578X1300801 ◽  
Author(s):  
Arvind Dabhade ◽  
Priti Patel ◽  
Ulhas Patil
Keyword(s):  
Antibacterial Activity ◽  
Protease Inhibitor ◽  
Deae Cellulose ◽  
Gel Permeation Chromatography ◽  
Apparent Molecular Weight ◽  
Exchange Chromatography ◽  
Lawsonia Inermis ◽  
Sds Page ◽  
Wide Range ◽  
And Staphylococcus Aureus

A thermo-stable, proteinaceous protease inhibitor (LPI) from Lawsonia inermis is reported. The LPI was purified from Lawsonia inermis seeds by subsequent ammonium sulfate precipitation, ion exchange chromatography (DEAE-Cellulose) and gel permeation chromatography (Sephadex-50). The purified protease inhibitor is effective against a wide range of proteases viz. papain, trypsin, pepsin and metallo-protease. The apparent molecular weight of the protease inhibitor is 19 kDa, determined by SDS-PAGE electrophoresis. The protease inhibitor was found to be stable at 70 °C for 30 min. It was also examined for antibacterial activity against Pseudomonas aeruginosa MTCC 7926 and Staphylococcus aureus NCIM 2079; the IC50 values of the purified LPI were 11.4 μg/mL and 16.6 μg/mL respectively.

Purification and characterization of exocellular proteases produced by a clinical isolate and a laboratory strain of Pseudomonas aeruginosa

1980 ◽  
Vol 26 (1) ◽  
pp. 77-86 ◽  
Author(s):  
S. E. Jensen ◽  
L. Phillippe ◽  
J. Teng Tseng ◽  
G. W. Stemke ◽  
J. N. Campbell
Keyword(s):  
Molecular Weight ◽  
Pseudomonas Aeruginosa ◽  
Clinical Isolate ◽  
Protease Activity ◽  
Gel Filtration ◽  
Laboratory Strain ◽  
Exchange Chromatography ◽  
The Other ◽  
Protease Production ◽  
Peptide Bonds

Exocellular protease production was examined in two separate strains of Pseudomonas aeruginosa, one a clinical isolate and the other a laboratory strain. Both strains produced two separate proteases (proteases 1 and 2) which were indistinguishable from one strain to the other. The two proteases were purified by a two-step procedure of gel filtration chromatography followed by ion-exchange chromatography. Proteases 1 and 2 were shown to be distinct serologically and unrelated by physicochemical parameters examined. Protease 1 was the major exocellular protein produced and contributed about 95% of the total protease activity of the culture. It was estimated to have a molecular weight of 34 850 and was also shown to contain 10% glucosamine by weight. Protease 2, in contrast, had an estimated molecular weight of 52750 and contained no detectable carbohydrate. Proteases 1 and 2 were both stimulated by Ca2+, and Mg2+ and inhibited by Co2+Zn2+, and 1,10-o-phenanthroline. Protease 1 was also inhibited by EDTA. In addition to protease activity, both proteases 1 and 2 demonstrated elastase activity as well as a limited collagenase activity. Specificity of the two proteases against synthetic peptides was, however, quite different. Protease 1, but not protease 2, showed a preference for peptide bonds in which the amino group was contributed by an amino acid with a hydrophobic R group.

scholarly journals HEMICELLULOSE APPARENT MOLECULAR SIZE CHANGES DURING SOFTENING OF PEACH FRUIT

HortScience ◽  
1994 ◽  
Vol 29 (7) ◽  
pp. 737d-737
Author(s):  
Supreetha Hegde ◽  
Niels Maness
Keyword(s):  
Molecular Weight ◽  
Cell Wall ◽  
Apparent Molecular Weight ◽  
Weight Fraction ◽  
Exchange Chromatography ◽  
Size Exclusion ◽  
Neutral Sugar ◽  
Fruit Softening ◽  
Peach Fruit ◽  
Cell Wall Polymers

Peach fruit softening appears to be associated with changes in cell wall polymers, particularly pectins and hemicelluloses. To determine changes of cell wall polymers associated with peach fruit softening, we conducted sequential extractions of pectin and hemicellulose from softening fruit. A more tightly bound hemicellulose fraction contained considerable amounts of pectin associated sugars. This fraction was separated into charged and neutral fractions, using anion exchange chromatography, and then fractionated into two apparent molecular weight classes by size exclusion chromatography. Virtually all of the charged fraction eluted in the higher apparent molecular weight fraction. The neutral sugar fraction segregated into both apparent molecular weight size classes, with a redistribution from the large to the small size class during softening. This redistribution was accompanied by changes in neutral sugar composition. A possible relationship between changes in this fraction and fruit softening will be discussed. Supported by USDA grant 92-34150-7190 and the Oklahoma Agricultural Experiment Station.

scholarly journals Macromolecules stimulating human granulocytic colony-forming cells, precursors of these cells, and primitive erythroid progenitors: some apparent nonidentities

Blood ◽  
1983 ◽  
Vol 61 (5) ◽  
pp. 960-966 ◽  
Author(s):  
T Hoang ◽  
NN Iscove ◽  
N Odartchenko
Keyword(s):  
Molecular Weight ◽  
Conditioned Medium ◽  
Gel Filtration ◽  
Apparent Molecular Weight ◽  
The Other ◽  
Erythroid Progenitors ◽  
Granulocytic Colony ◽  
Ph Range ◽  
Direct Primary ◽  
The Relationship

Abstract The relationship between molecules having granulocyte colony- stimulating activity (G-CSA), erythroid burst-promoting activity (E- BPA), and activity promoting increase in the number of granulocytic progenitors in liquid culture (delta GPA) was explored in conditioned medium from human leukocytes (HLCM) and human placenta (HPCM). As tested on human hemopoietic progenitors in culture, G-CSA eluted from Sephadex G100 as a single peak with apparent molecular weight of 25,000, separating partially from E-BPA and delta GPA, which both had an apparent molecular weight of 45,000. All three activities eluted together from hydroxyapatite at low molarity phosphate. Their charge properties were also similar and all three electrofocused in flat gel beds in the pH range near 5.4. On both hydroxyapatite and isoelectric focusing, delta GPA sometimes separated partially from the other two activities but not consistently. The gel filtration result shows that in conditioned medium of human origin, molecules having G-CSA are not the same as those having delta GPA, suggesting a dual factor requirement in the granulocytic lineage reminiscent of that in the erythroid pathway. The results suggesting that delta GPA might differ from E-BPA, on the other hand, were not consistent enough to establish their nonidentity. Single micromanipulated cells proved capable of forming erythroid or granulocytic colonies in the presence of either crude or partially purified activity. The results establish that human colony-forming cells are direct primary targets of growth factors in HLCM and HPCM.

Characterisation of a Neutral Protease Produced by Micrococcus luteus

1996 ◽  
Vol 51 (5-6) ◽  
pp. 429-431 ◽  
Author(s):  
M.O. Ilori ◽  
O.O. Amund ◽  
O. Omidiji
Keyword(s):  
Molecular Weight ◽  
Citric Acid ◽  
Proteolytic Enzyme ◽  
Micrococcus Luteus ◽  
Gel Filtration ◽  
Apparent Molecular Weight ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Optimum Temperature ◽  
Optimum Ph

Abstract A proteolytic enzyme produced by a cassava-ferment­ing strain of Micrococcus luteus was extracted and puri­fied 50-fold by gel filtration and ion exchange chromatography. The optimum pH for the enzyme was 7.0, the optimum temperature 25 °C, the apparent molecular weight 42 kDa and the Km value, 0.45 mg ml-1 with casein as substrate. The enzyme was stimulated by Ca2+ and Mg2+ but inhibited by Zn2+ and Co2+ ions. Other inhibitors were EDTA, KCN, citric acid and L-cysteine indicating the enzyme to be a metalloprotease.

Zur Strahleninaktivierung von Ribonuclease

1966 ◽  
Vol 21 (3) ◽  
pp. 224-231 ◽  
Author(s):  
Horst Jung ◽  
Helga Schüssler
Keyword(s):  
Molecular Weight ◽  
Gamma Radiation ◽  
Enzymatic Activity ◽  
Column Chromatography ◽  
Apparent Molecular Weight ◽  
Ribonuclease A ◽  
Nitrogen Atmosphere ◽  
The Other ◽  
Radiation Action ◽  
Higher Doses

Ribonuclease A was irradiated in water with 60Co gamma radiation, and the products formed were separated according to their molecular weight by column chromatography on Sephadex G-50. When the irradiations are carried out in nitrogen atmosphere aggregation to dimers and higher polymers is observed. An appreciable fraction of this aggregated component retains enzymatic activity. At higher doses the enzymatic activity of the aggregates is inactivated at the same rate as monomer ribonuclease A. Irradiation in air yields two components; one is equivalent to that found in nitrogen atmosphere, the other has an apparent molecular weight of about 20 000 but contains no enzymatic activity. This last component is not observed when methionine is present during irradiation. In methionine containing solutions all inactivited ribonuclease molecules exist in the form of dimers and polymers. Irradiation in the dry state leads to the same result. Consequently, every model designed to describe the radiation action on ribonuclease has to consider the fact that in solution, in the presence of a protecting agent, and in the dry state the loss of enzymatic activity is always accompanied by aggregation to products with increased molecular weight.

scholarly journals The N-acetylhexosaminidase components of the ram testis and epididymis

1973 ◽  
Vol 133 (3) ◽  
pp. 593-599 ◽  
Author(s):  
Sarah Bullock ◽  
Bryan Winchester
Keyword(s):  
Molecular Weight ◽  
Ion Exchange ◽  
Isoelectric Focusing ◽  
Catalytic Properties ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Multiple Forms ◽  
The Individual

Three and four N-acetylhexosaminidase components, from ram testis and epididymis respectively, have been separated by ion-exchange chromatography on DEAE-cellulose. Although they all have the same molecular weight (approx. 140000) and very similar catalytic properties towards the synthetic substrates, 4-methylumbelliferyl N-acetyl-β-glucosaminide and N-acetyl-β-galactosaminide, isoelectric focusing of the individual components showed that each had a distinct pI value. Isoelectric focusing has also been used to demonstrate the occurrence of multiple forms in ejaculated ram semen.

On the structure alteration of crosslinkable gelatin coupled with methacrylic acid and its hydrogel

e-Polymers ◽  
2012 ◽  
Vol 12 (1) ◽  
Author(s):  
Xiaohong Hu ◽  
Dan Li ◽  
Feng Zhou ◽  
Changyou Gao
Keyword(s):  
Molecular Weight ◽  
Triple Helix ◽  
Sol Gel ◽  
Average Molecular Weight ◽  
Gel Permeation Chromatography ◽  
Apparent Molecular Weight ◽  
Small Range ◽  
Scanning Calorimetry ◽  
Physical Chemical ◽  
Substitution Degree

AbstractPhysical structures of a crosslinkable gelatin derivative (GM) were studied in terms of alteration of apparent molecular weight, triple helix content and mechanical strength. The GM with a substitution degree (DS) of 49% and 79% was prepared by grafting mechacrylic acid (MA), which was able to form injectable hydrogel by photoinitiating polymerization. The zeta potential was increased along the increase of DS. After modification, the apparent number-average molecular weight (Mn) detected by gel permeation chromatography was decreased to about 2/3 of gelatin, while the apparent weight-average molecular weight (Mw) was changed within a small range. Differential scanning calorimetry and circular dichroism (CD) revealed that ability of triple-helix formation of GM was decreased along with the increase of DS and decrease of GM concentration. After photocrosslinking, the sol-gel transition of GM49 physical-chemical hydrogel still existed, but completely disappeared for its chemical hydrogel. The physical-chemical hydrogel showed a larger storage modulus at 20°C than at 37°C as a result of additional physical crosslinking.

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