scholarly journals The Effect of Temperature on the Chromatography of Insulin on Deae-Cellulose

1960 ◽  
Vol 13 (1) ◽  
pp. 69 ◽  
Author(s):  
IJ O'donnell ◽  
EOP Thompson
Keyword(s):  
Ionic Strength ◽  
Elution Curve ◽  
Deae Cellulose ◽  
Minor Component ◽  
Ammonia Removal ◽  
Effect Of Temperature ◽  
Bovine Plasma ◽  
Three Samples ◽  
Ph Range ◽  
A Minor

The effect of ionic strength (range 0,15-0, 3), pH (range 7-9), and temperature (range I-25�C) on the chromatographic behaviour of three samples of insulin on diethylaminoethyl (DEAE)-cellulose columns has been studied. These three factors have a similar effect, a decrease of temperature or pH and an increase in ionic strength lowering the elution volume of the protein. The marked effect of temperature is not due to aggregation-disaggregation of the insulin since bovine plasma albumin which does not aggregate reversibly also showed this effect. The desamido component of insulin could not be detected in commercial insulin under the conditions studied but a minor component varying from 2-6 per cent. of the insulin was separated, as well as various amounts of bound ammonia. Removal of zinc from the insulin did not affect the elution curve.

scholarly journals Structural defects in rat liver deoxyribonucleic acid. Endogenous single-strained regions in comparison with damage induced in vivo by a carcinogen

1979 ◽  
Vol 179 (2) ◽  
pp. 341-352 ◽  
Author(s):  
B W Stewart ◽  
P H Huang ◽  
M J Brian
Keyword(s):  
Rat Liver ◽  
Structural Damage ◽  
Deae Cellulose ◽  
Minor Component ◽  
Structural Defects ◽  
Structural Abnormality ◽  
Denaturation Kinetics ◽  
Limited Digestion ◽  
A Minor

Rat liver DNA may be separated into two fractions by stepwise elution from benzoylated-DEAE-cellulose with NaCl and caffeine solutions respectively. Other studies using bacterical and yeast DNA suggested that the first fraction contains native DNA, whereas the second may exhibit some degree of single-stranded character. In the present experiments, chromatography of DNA was monitored by labelling in vivo with [methyl-3H]thymidine in rats previously subjected to partial hepatectomy. In animals killed up to 1 h after thymidine injection, radioactivity eluted in the second fraction was inversely related to the incorporation time, being greatest when animals were killed 10 min after radioisotope injection. However, for most experiments, animals were allowed to survive 2-4 weeks after surgery before use, analysis being made on non-dividing DNA. Under these conditions, the proportion of caffeine-eluted DNA was decreased by subjecting the preparation to shear, before chromatography. A procedure that resulted in 12% of the recovered radioactivity being eluted with caffeine was adopted for experiments involving comparisons of the two DNA fractions. Under these conditions, cross-contamination could be detected by rechromatography, but this did not preclude distinction being made between the two fractions in terms of DNA structure. NaCl-eluted DNA did not bind to nitrocellulose filters. Caffeine-eluted DNA was retained by the filters and released by washing with 3mM-Tris/HCl, pH9.4. The fractions did not differ in terms of isopycnic centrifugation in CsCl. The NaCl-eluted fraction migrated as a single band in polyacrylamide gels, and this pattern was not modified by prior digestion with Neurospora crassa endonuclease. In contrast, caffeine-eluted DNA contained a minor component having a wide molecular-weight distribution and was subject to limited digestion by the endonuclease. The kinetics of denaturation of NaCi-eluted DNA in the presence of formaldehyde, in common with unfractionated DNA, were consistent with double-stranded structure. The same analysis of caffeine-eluted DNA revealed structural abnormality equivalent to two defects per 10000 base-pairs. The data are consistent with the minor fraction of rat liver DNA, separated by using benzoylated-DEAE-cellulose, containing regions of local denaturation. We previously showed that administration of the hepatocarcinogen dimethylnitrosamine is associated with an increase in the proportion of caffeine-eluted DNA. In terms of most analysis, differences between DNA fraction from nitrosamine-treated rats were similar to differences exhibited by preparations from control animals. However, structural analysis using denaturation kinetics indicated defects in both the NaCl- and caffeine-eluted DNA isolated from nitrosamine-treated rats. The two fractions differed from each other in that caffeine-eluted DNA exhibited a degree of structural damage far greater than that detected in any preparation from control animals...

scholarly journals Outstanding Characteristics of Thrombokinase Isolated from Bovine Plasma

1963 ◽  
Vol 47 (2) ◽  
pp. 315-327 ◽  
Author(s):  
J. H. Milstone ◽  
N. Oulianoff ◽  
V. K. Milstone
Keyword(s):  
Methyl Ester ◽  
Continuous Flow ◽  
Deae Cellulose ◽  
Stability Curve ◽  
Bovine Plasma ◽  
Rapid Production ◽  
The Stability ◽  
Ph Range ◽  
Single Boundary ◽  
Electrophoretic Fraction

Thrombokinase has been isolated from bovine plasma by a procedure which begins with the highly purified product of a previously described method, chromatographs it on DEAE-cellulose, and then fractionates it by continuous flow electrophoresis, yielding 0.2 mg per liter of oxalated plasma. The electrophoretic fraction has shown a single boundary in the ultracentrifuge; and its esterase activity on toluenesulfonylarginine methyl ester has been about the same as that of thrombokinase previously isolated by repeated electrophoretic fractionations. Thrombokinase is a euglobulin with minimum solubility near pH 5.0. It is most stable within the pH range 7.5 to 9.5; but there is also a peak in the stability curve near pH 1.8. A few micrograms of thrombokinase per milliliter can activate prothrombin in the presence of EDTA. A few thousandths of a microgram causes rapid production of thrombin in the system: prothrombin, thrombokinase, calcium chloride, phosphatide, "accelerator." But, thrombokinase has less than 1/175 the proteolytic activity of crystallized trypsin.

scholarly journals N-Acetyl-β-d-hexosaminidase component A. Different forms in human tissues and fluids

1973 ◽  
Vol 135 (3) ◽  
pp. 457-462 ◽  
Author(s):  
J. U. Ikonne ◽  
R. B. Ellis
Keyword(s):  
Salt Concentration ◽  
Human Liver ◽  
Deae Cellulose ◽  
Minor Component ◽  
Human Tissues ◽  
Serum Enzyme ◽  
Hexosaminidase A ◽  
Minor Form ◽  
A Minor ◽  
Cellulose Column

1. Hexosaminidase A of human serum was resolved into two components, a minor form with properties identical with those of the single hexosaminidase A component of human liver, and a major form with significantly different properties. 2. The major serum hexosaminidase A form was eluted from a DEAE-cellulose column at a lower salt concentration than that required to elute the liver form. 3. A multiple-pass technique was used to elute the major serum enzyme A from a Sephadex G-150 column before that of liver enzyme A. 4. Clostridium perfringens neuraminidase converted the major component of serum hexosaminidase A into a form that was held less tightly by DEAE-cellulose, but the minor component of the A enzyme of serum, and the A enzyme of liver were not affected. 5. The hexosaminidase A from tears was similar to the A enzyme from serum, whereas those from several human tissues and from urine and lymph were similar to the liver form. 6. The A enzyme from serum may be derived from the A enzyme from liver by glycosylation before secretion.

scholarly journals Studies on the high-sulphur proteins of reduced Merino wool. Amino acid sequence of protein SCMKB-IIIA3

1973 ◽  
Vol 133 (4) ◽  
pp. 641-654 ◽  
Author(s):  
Louis S. Swart ◽  
Thomas Haylett
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Minor Component ◽  
Amino Acid Sequences ◽  
Edman Degradation ◽  
Merino Wool ◽  
C Terminus ◽  
A Minor

The complete amino acid sequences of wool protein SCMKB-IIIA3 (131 residues) and a minor component SCMKB-IIIA3A (130 residues) have been determined. The proteins are mutually homologous and have free threonine as the N-terminal residue and carboxymethylcysteine as the C-terminus. The peptides used for the sequence work were obtained by trypsin, thermolysin, pepsin and chymotrypsin digestions and were fractionated by chromatography on DEAE-cellulose, gel filtration on Sephadex G-25 and G-50, paper chromatography and electrophoresis. The Edman degradation method (employing both the Beckman Sequencer and a non-automatic procedure) was used to obtain the sequences of the peptides.

Georgenia sediminis sp. nov., a moderately thermophilic actinobacterium isolated from sediment

2013 ◽  
Vol 63 (Pt_11) ◽  
pp. 4243-4247 ◽  
Author(s):  
Zhi-Qing You ◽  
Jie Li ◽  
Sheng Qin ◽  
Xin-Peng Tian ◽  
Fa-Zuo Wang ◽  
...  
Keyword(s):  
Type Species ◽  
Minor Component ◽  
Optimal Growth ◽  
Rrna Gene ◽  
Content Type ◽  
Moderately Thermophilic ◽  
Link Type ◽  
Nacl Concentration ◽  
Ph Range ◽  
A Minor

A Gram-stain-positive actinobacterium, designated strain SCSIO 15020T, was isolated from sediment of the South China Sea, and characterized by using a polyphasic approach. The temperature range for growth was 24–60 °C, with optimal growth occurring at 50 °C. The pH range for growth was 6–10 (optimum pH 8–9). The NaCl concentration range for growth was 0–5 % (w/v). The peptidoglycan type was A4α. Polar lipids contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside and an unknown polar lipid. The major menaquinone was MK-8(H4); MK-7(H4) was present as a minor component. The major fatty acids (>5 %) were anteiso-C15 : 0, iso-C15 : 0 and iso-C16 : 0. The DNA G+C content of strain SCSIO 15020T was 73.2 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain SCSIO 15020T belonged to the genus Georgenia , with the closest neighbours being Georgenia muralis 1A-CT (96.3 % similarity), Georgenia thermotolerans TT02-04T (95.7 %) and Georgenia ruanii YIM 004T (95.6 %). Based on evidence from the present polyphasic study, strain SCSIO 15020T is considered to represent a novel species of the genus Georgenia , for which the name Georgenia sediminis sp. nov. is proposed. The type strain is SCSIO 15020T ( = DSM 25884T = NBRC 108941T).

scholarly journals Interdental Supragingival Plaque—A Natural Habitat of Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Campylobacter rectus, and Prevotella nigrescens

1994 ◽  
Vol 73 (8) ◽  
pp. 1421-1428 ◽  
Author(s):  
R. Gmur ◽  
B. Guggenheim
Keyword(s):  
Periodontal Diseases ◽  
Natural Habitat ◽  
Minor Component ◽  
Actinobacillus Actinomycetemcomitans ◽  
Periodontal Pathogens ◽  
Campylobacter Rectus ◽  
Prevotella Nigrescens ◽  
Supragingival Plaque ◽  
Three Samples ◽  
A Minor

The aim of this study was to test the hypothesis that suspected periodontal pathogens form a minor component of the supragingival plaque of individuals without periodontal diseases. Twenty-one dental hygienist trainees with a mean age of 23.5 years were twice sampled for interdental plaque between 1st and 2nd molars in all quadrants. The samples were assessed for Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Campylobacter rectus, Porphyromonas gingivalis, and the Prevotella intermedia/Prevotella nigrescens group of organisms. Bacteria of this group were predominantly P. nigrescens and showed both the highest prevalence (100%) and the highest colonization density of the investigated species. Seven of 21 samples harbored A. actinomycetemcomitans. Serotypes a, b, and c were found in three samples each, while serotype e was present in one sample. Three subjects had two different A. actinomycetemcomitans serotypes. Bacteroides forsythus and C. rectus were detected in 10 (48%) and nine (43%) subjects, respectively. The detected cell numbers accounted for approximately 0.01% to 1% of the sampled flora. In contrast, P. gingivalis was found only in a single sample, which in addition harbored B. forsythus, C. rectus, A. actinomycetemcomitans (serotypes b and e), and P. intermedia. These results suggest that the investigated periodontal bacteria are not "exogenous pathogens", but amphibiotic, opportunistic microorganisms which may have a natural habitat in the supragingival plaque of the interproximal area of molars.

scholarly journals Identification of β-N-acetylhexosaminidase A in mouse tissues with the fluorigenic substrate 4-methylumbelliferyl-β-N-acetylglucosamine 6-sulphate

1988 ◽  
Vol 252 (2) ◽  
pp. 617-620 ◽  
Author(s):  
T Beccari ◽  
A Orlacchio ◽  
J L Stirling
Keyword(s):  
Mouse Liver ◽  
Specific Activity ◽  
Deae Cellulose ◽  
Minor Component ◽  
Mouse Tissue ◽  
Intermediate Form ◽  
Mouse Tissues ◽  
A Minor

beta-N-Acetylhexosaminidase from mouse tissue was separated into its constituent isoenzymes on DEAE-cellulose and its activity was monitored with 4-methylumbelliferyl-beta-N-acetylglucosamine and 4-methylumbelliferyl-beta-N-acetylglucosamine 6-sulphate. Forms corresponding to the human isoenzymes A (acidic), B (basic) and an ‘intermediate’ form were present in mouse liver and spleen, whereas in kidney the B and ‘intermediate’ forms predominated, with A present only as a minor component. In brain the ‘intermediate’, A and C activities were detected. Testis had predominantly A activity, whereas epididymis, the tissue with the highest specific activity of beta-N-acetylhexosaminidase, had an abundance of the ‘intermediate’ form, but was almost entirely lacking in the A form.

Methods for the Concentration of Bovine Ac-Globulin (Factor V)

1961 ◽  
Vol 06 (03) ◽  
pp. 435-444 ◽  
Author(s):  
Ricardo H. Landaburu ◽  
Walter H. Seegers
Keyword(s):  
Room Temperature ◽  
Barium Carbonate ◽  
Deae Cellulose ◽  
Reducing Agents ◽  
Factor V ◽  
Isoelectric Precipitation ◽  
Stabilizing Agent ◽  
Bovine Plasma ◽  
The Stability

SummaryAn attempt was made to obtain Ac-globulin from bovine plasma. The concentrates contain mostly protein, and phosphorus is also present. The stability characteristics vary from one preparation to another, but in general there was no loss before 1 month in a deep freeze or before 1 week in an icebox, or before 5 hours at room temperature. Reducing agents destroy the activity rapidly. S-acetylmercaptosuccinic anhydride is an effective stabilizing agent. Greatest stability was at pH 6.0.In the purification bovine plasma is adsorbed with barium carbonate and diluted 6-fold with water. Protein is removed at pH 6.0 and the Ac-globulin is precipitated at pH 5.0. Rivanol and alcohol fractionation is followed by chromatography on Amberlite IRC-50 or DEAE-cellulose. The final product is obtained by isoelectric precipitation.

THE RELATIVE DISTRIBUTION OF THYROXINE, TRIIODOTHYRONINE AND 3,3′,5′-(REVERSE)-TRIIODOTHYRONINE IN VARIOUS FRACTIONS OF THYROGLOBULIN

1978 ◽  
Vol 88 (2) ◽  
pp. 298-305 ◽  
Author(s):  
Peter Laurberg
Keyword(s):  
Ionic Strength ◽  
Guinea Pig ◽  
Sucrose Gradient ◽  
Deae Cellulose ◽  
High Ionic Strength ◽  
Relative Distribution ◽  
Thyroid Extract ◽  
Reverse Triiodothyronine ◽  
Thyroid Secretion ◽  
Stable Iodine

ABSTRACT Thyroglobulin fractions rich and poor in new thyroglobulin were separated by means of DEAE-cellulose chromatography of dog thyroid extracts and by zonal ultracentrifugation in a sucrose gradient of guinea pig thyroid extract incubated at low temperature. The distribution of thyroxine, triiodothyronine and 3,3′,5′-(reverse)-triiodothyronine in hydrolysates of the different fractions was estimated by radioimmunoassays. Following DEAE-cellulose chromatography there was a small but statistically significant increase in the T4/T3 ratio in thyroglobulin fractions eluted at high ionic strength - that is fractions relatively rich in stable iodine but poor in fresh thyroglobulin. There were no differences in the T4/rT3 ratios between the different fractions. The ratios between iodothyronines were almost identical in the various thyroglobulin fractions following zonal ultracentrifugation in a sucrose gradient of cold treated guinea pig thyroid extract. These findings lend no support to the possibility that a relatively high content of triiodothyronines in freshly synthesized thyroglobulin modulates the thyroid secretion towards a preferential secretion of triiodothyronine and 3,3′,5′-(reverse)-triiodothyronine at the expense of the secretion of thyroxine.

Protonation effects on dynamic flux properties of aqueous metal complexes

2009 ◽  
Vol 74 (10) ◽  
pp. 1543-1557 ◽  
Author(s):  
Herman P. Van Leeuwen ◽  
Raewyn M. Town
Keyword(s):  
Complex Formation ◽  
Metal Ion ◽  
Good Description ◽  
Minor Component ◽  
Outer Sphere ◽  
Metal Species ◽  
Central Metal ◽  
Inner Sphere ◽  
A Minor ◽  
Protonated Ligand

The degree of (de)protonation of aqueous metal species has significant consequences for the kinetics of complex formation/dissociation. All protonated forms of both the ligand and the hydrated central metal ion contribute to the rate of complex formation to an extent weighted by the pertaining outer-sphere stabilities. Likewise, the lifetime of the uncomplexed metal is determined by all the various protonated ligand species. Therefore, the interfacial reaction layer thickness, μ, and the ensuing kinetic flux, Jkin, are more involved than in the conventional case. All inner-sphere complexes contribute to the overall rate of dissociation, as weighted by their respective rate constants for dissociation, kd. The presence of inner-sphere deprotonated H2O, or of outer-sphere protonated ligand, generally has a great impact on kd of the inner-sphere complex. Consequently, the overall flux can be dominated by a species that is a minor component of the bulk speciation. The concepts are shown to provide a good description of experimental stripping chronopotentiometric data for several protonated metal–ligand systems.

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