Effect of diet and drugs on the qualitative and quantitative distribution of cytochromes P-450 in rat liver

1980 ◽  
Vol 58 (4) ◽  
pp. 331-335 ◽  
Author(s):  
Daniel S. Sitar ◽  
Ellen R. Gordon
Keyword(s):  
Ion Exchange ◽  
Control Diet ◽  
Caloric Intake ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Sprague Dawley ◽  
Quantitative Distribution ◽  
Qualitative And Quantitative ◽  
Purina Chow ◽  
Sprague Dawley Rats

Male Sprague–Dawley rats were fed either Purina chow or a nutritionally adequate liquid control diet containing either 4 or 38% of the total caloric intake as fat. The hepatic cytochromes P-450 were induced in these animals either by the administration of phenobarbital, β-naphthoflavone, or isocaloric replacement of the carbohydrate of the liquid diets by ethanol at a level of 36% of total calories for 4 or 6 weeks. Three distinct groups of cytochromes P-450 could be efficiently separated by ion-exchange chromatography of microsomal preparations from these rats. The concentration of each group of cytochromes P-450 was markedly affected by variations in lipid and carbohydrate content of the diet as well as by administration of drugs.

β-Glucuronidase Isoenzymes of Rat-Liver Lysosomes

1973 ◽  
Vol 51 (7) ◽  
pp. 973-979 ◽  
Author(s):  
M. Potier ◽  
R. Gianetto
Keyword(s):  
Gel Filtration ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Sprague Dawley ◽  
Active Components ◽  
Precipitation Treatment ◽  
Sprague Dawley Rats ◽  
Liver Lysosomes ◽  
Rat Liver Lysosomes

β-Glucuronidase (β-D-glucuronide glucuronohydrolase, EC 3.2.1.31) from liver lysosomes of male Sprague–Dawley rats was purified by (NH4)2SO4 precipitation, treatment with trypsin and gel filtration on Sephadex G-200, and then fractionated into five active components by ion-exchange chromatography on DEAE-Sephadex A-25 or DEAE-cellulose. All five isoenzymes were shown to be electrophoretically and ultracentrifugally homogeneous. Their sedimentation coefficients range from 12.27 to 13.39. All five isoenzymes appear to be glycoproteins.

Abstract 046: High Fructose Intake Increases Phosphorylation and Trafficking of the Na/K/2Cl Cotransporter (NKCC22) In Rat Thick Ascending Limbs

Hypertension ◽  
2014 ◽  
Vol 64 (suppl_1) ◽  
Author(s):  
Gustavo R Ares ◽  
Mohammed Z Haque ◽  
Pablo A Ortiz
Keyword(s):  
Drinking Water ◽  
Apical Membrane ◽  
Control Diet ◽  
Caloric Intake ◽  
Thick Ascending Limb ◽  
Sprague Dawley ◽  
Added Sugars ◽  
Sprague Dawley Rats ◽  
Enhanced Expression ◽  
Salt Sensitive Hypertension

A third of the US population consumes 20-40% of their caloric intake from added sugars with half of those calories from fructose. Fructose consumption is linked to salt-sensitive hypertension in humans and rodents. We found that feeding Sprague-Dawley rats 20% fructose in their drinking water did not increase blood pressure unless a high salt diet was added. The thick ascending limb (TAL) reabsorbs 25% of filtered NaCl, primarily via NKCC2. NKCC2 activity is increased by enhanced expression at the apical membrane and phosphorylation at Thr 96/101 . We found enhanced NKCC2 activity and trafficking in genetic models of salt-sensitive hypertension. However, the effect of fructose on NKCC2 regulation is unknown. Thus, we hypothesized that a fructose enriched diet stimulates NKCC2 activity. Sprague-Dawley rats were fed control diet or 20% fructose in drinking water for 1 week. TALs were isolated and phosphorylated and total NKCC2 was measured by surface biotinylation followed by western blot. A fructose-enriched diet increased surface-to-intracellular NKCC2 ratio by 60 ± 23% ( p< 0.05) and increased NKCC2 phosphorylation at Thr 96/101 by 8.02 ± 2.67 fold ( p <0.05). NKCC2 phosphorylation at the plasma membrane was also increased by 4.5 fold ( p <0.05). Total NKCC2 expression was reduced by 40.1 ± 8.9% ( p< 0.05). Since phosphorylation of NKCC2 at Thr 96/101 is mediated by STE20- and SPS1-related proline and alanine-rich kinases (SPAK) and oxidative stress-responsive kinase 1 (OSR1), we studied whether fructose stimulates expression and/or activity of SPAK/OSR1. Total SPAK/OSR1 expression was not enhanced by one week of a fructose diet. However, phosphorylation at Ser373 was enhanced by 2.8 ± 0.3 fold ( p< 0.05). We concluded that a fructose enriched diet increases phosphorylation and trafficking of NKCC2, enhancing its accumulation at the apical membrane. Moreover, a fructose diet increased SPAK/OSR1 kinases phosphorylation, suggesting they may be responsible for the enhanced NKCC2 phosphorylation. Our data suggest that a fructose-enriched diet promotes salt-sensitive hypertension in part by stimulating NKCC2 and TAL-dependent NaCl reabsorption.

Studies on an Inhibitor of Plasminogen Activation in Human Serum

1973 ◽  
Vol 30 (02) ◽  
pp. 414-424 ◽  
Author(s):  
Ulla Hedner
Keyword(s):  
Ion Exchange ◽  
Human Serum ◽  
Antithrombin Iii ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Specific Adsorption ◽  
Partial Purification ◽  
Plasminogen Activation ◽  
Preparative Electrophoresis

SummaryA procedure is described for partial purification of an inhibitor of the activation of plasminogen by urokinase and streptokinase. The method involves specific adsorption of contammants, ion-exchange chromatography on DEAE-Sephadex, gel filtration on Sephadex G-200 and preparative electrophoresis. The inhibitor fraction contained no antiplasmin, no plasminogen, no α1-antitrypsin, no antithrombin-III and was shown not to be α2 M or inter-α-inhibitor. It contained traces of prothrombin and cerulo-plasmin. An antiserum against the inhibitor fraction capable of neutralising the inhibitor in serum was raised in rabbits.

Review on the Application of Mixed-mode Chromatography for Separation of Structure Isoforms

2018 ◽  
Vol 20 (1) ◽  
pp. 56-60 ◽  
Author(s):  
Tsutomu Arakawa
Keyword(s):  
Ion Exchange ◽  
Mixed Mode ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Charged State ◽  
Partial Structure ◽  
Reverse Phase ◽  
Oligomer Formation ◽  
Structure Rearrangement ◽  
Disulfide Scrambling

Proteins often generate structure isoforms naturally or artificially due to, for example, different glycosylation, disulfide scrambling, partial structure rearrangement, oligomer formation or chemical modification. The isoform formations are normally accompanied by alterations in charged state or hydrophobicity. Thus, isoforms can be fractionated by reverse-phase, hydrophobic interaction or ion exchange chromatography. We have applied mixed-mode chromatography for fractionation of isoforms for several model proteins and observed that cation exchange Capto MMC and anion exchange Capto adhere columns are effective in separating conformational isoforms and self-associated oligomers.

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