Studies on Acute Phase Proteins of Rat Serum. III. Site of Synthesis of Albumin and α1-Acid Glycoprotein and the Contents of These Proteins in Liver Microsome Fractions from Rats Suffering from Induced Inflammation

1973 ◽  
Vol 51 (7) ◽  
pp. 1034-1045 ◽  
Author(s):  
J. C. Jamieson ◽  
F. E. Ashton
Keyword(s):  
Liver Microsome ◽  
Acute Phase Proteins ◽  
Specific Radioactivity ◽  
Rat Serum ◽  
Experimental Animals ◽  
Acid Glycoprotein ◽  
Inflammatory Agent ◽  
Microsome Fraction ◽  
Rapid Stimulation ◽  
Short Times

Following injection of L-ieucine-3H or D-glucosamine-14C to normal rats or rats suffering from inflammation for 12 h there was a delay of 10–15 min before label appeared in albumin and α1-acid glycoprotein in serum; thereafter, there were rapid increases in specific radioactivities. The specific radioactivity of L-leucine-3H in albumin in serum from normal and experimental animals was not significantly different; however, there was a more rapid increase in specific radioactivities of both labelled compounds in α1-acid glycoprotein isolated from serum from experimental animals. Immunological techniques coupled with radioautography indicated that the microsome fraction of liver was the subcellular site of synthesis of albumin and of carbohydrate and polypeptide moieties of α1-acid glycoprotein in normal and experimental animals. The contents of albumin and α1-acid glycoprotein in liver microsome fractions from normal and experimental animals were determined by application of the quantitative precipitin technique to Lubrol-W extracts of microsome material. Little change in content of albumin associated with microsome material was found as a result of inflammation; however, there was a significant increase in the content of α1-acid glycoprotein in microsome material from experimental animals, reaching a maximum at 8–12 h after inflammation. On the basis of these latter results, it is suggested (1) that there is a fairly rapid stimulation of synthesis of α1-acid glycoprotein by liver in response to inflammation and (2) that the increased content of α1-acid glycoprotein associated with microsome material at short times of exposure to inflammatory agent is responsible for the increased content of this protein found in serum at longer times of exposure to inflammatory agent.

Studies on Acute Phase Proteins of Rat Serum. II. Determination of the Contents of α1-Acid Glycoprotein, α2-Macroglobulin, and Albumin in Serum from Rats Suffering from Induced Inflammation

1972 ◽  
Vol 50 (8) ◽  
pp. 871-880 ◽  
Author(s):  
J. C. Jamieson ◽  
F. E. Ashton ◽  
A. D. Friesen ◽  
B. Chou
Keyword(s):  
Acute Phase ◽  
Acute Phase Proteins ◽  
Rat Serum ◽  
Experimental Animals ◽  
Acid Glycoprotein ◽  
Inflammatory Agent ◽  
Glycoprotein Biosynthesis ◽  
Α2 Macroglobulin

A quantitative precipitin technique has been employed to determine the contents of α1-acid glycoprotein, α2-macroglobulin, and albumin in serum from control rats and rats suffering from induced inflammation for 5–96 h. There was an increase in the content of α1-acid glycoprotein and α2-macroglobulin in serum from experimental animals reaching a maximum at 48–72 h after administration of inflammatory agent indicating that both proteins are acute phase globulins. There was only a slight change in the content of albumin in serum from experimental animals when compared with controls. Studies involving incorporation of labelled precursors of glycoprotein biosynthesis into α1-acid glycoprotein and α2-macroglobulin indicated that the most likely explanation for the increase in α1-acid glycoprotein and α2-macroglobulin in serum from experimental animals was an increase in the rates of synthesis of the two proteins in question.

Studies on Acute Phase Proteins of Rat Serum. V. Effect of Induced Inflammation on the Synthesis of Albumin and α1-Acid Glycoprotein by Liver Slices

1975 ◽  
Vol 53 (4) ◽  
pp. 401-414 ◽  
Author(s):  
J. C. Jamieson ◽  
K. E. Morrison ◽  
D. Molasky ◽  
B. Turchen
Keyword(s):  
Acute Phase ◽  
Polypeptide Chain ◽  
Serum Proteins ◽  
Acute Phase Proteins ◽  
Rat Serum ◽  
Acid Glycoprotein ◽  
Liver Slices ◽  
Microsome Fraction ◽  
Lower Capacity ◽  
Acute Phase Serum

Liver slices from normal rats and those suffering from inflammation for 24–48 h were incubated with L-[14C]leucine or D-[I4C]glucosamine. Immunological techniques coupled with radioautography indicated that the microsome fraction prepared from slices contained the subcellular site of synthesis of the polypeptide chain of serum albumin, and the polypeptide and carbohydrate chains of α1-acid glycoprotein; both proteins were also present in the medium in labelled forms. The contents of albumin and α1-acid glycoprotein in the medium and in extracts of liver from experiments with liver slices from control rats and 8–72 h experimental rats were determined using the quantitative precipitin technique. There was a net increase in synthesis of both proteins when slices from control and experimental animals were used, the increase showing up in medium proteins. However, slices from livers from 8–72 h experimental rats had a greater capacity for synthesis of α1-acid glycoprotein and a lower capacity for synthesis of albumin than slices from livers from control rats, the greatest changes occurring with slices from 24 h experimental rats. Changes in synthetic capacities of liver slices from experimental rats for albumin and α1-acid glycoprotein were always accompanied by large increases in specific radioactivities of total medium proteins when experiments involved incubation of slices with L-[3H]leucine and D-[14C]glucosamine. It is suggested that the increase in specific radioactivities of medium proteins following incubation of liver slices from experimental rats with labelled leucine and glucosamine is a characteristic of the response of liver to inflammation, and reflects changes in the capacity of liver for the synthesis of α1-acid glycoprotein and other acute phase serum proteins.

Studies on Acute Phase Proteins of Rat Serum. IV. Pathway of Secretion of Albumin and α1-Acid Glycoprotein from Liver

1973 ◽  
Vol 51 (9) ◽  
pp. 1281-1291 ◽  
Author(s):  
J. C. Jamieson ◽  
F. E. Ashton
Keyword(s):  
Endoplasmic Reticulum ◽  
Acute Phase ◽  
Rough Endoplasmic Reticulum ◽  
Golgi Complex ◽  
Acute Phase Proteins ◽  
Smooth Endoplasmic Reticulum ◽  
Rat Serum ◽  
Experimental Animals ◽  
Acid Glycoprotein ◽  
Ionic Detergent

Normal rats and those suffering from inflammation for 12 h were given pulse injections of L-leucine-3H and D-glucosamine-14C and were killed at 3–60 min thereafter. Rough and smooth microsome fractions and a Golgi-enriched fraction were prepared from liver and the fractions were extracted with the non-ionic detergent Lubrol-W. Albumin and acute phase α1-acid glycoprotein were isolated from the extracts by application of a quantitative precipitin technique employing antisera to albumin and α1-acid glycoprotein, and specific radioactivities were determined. The results indicated that the pathway of secretion of albumin and α1-acid glycoprotein was rough endoplasmic reticulum → smooth endoplasmic reticulum → Golgi complex → blood. Although there was little difference in the pathway or rate of secretion of the proteins under study in normal and experimental animals, there was an increase in specific radioactivities of labelled leucine and glucosamine associated with α1-acid glycoprotein when isolated from subcellular fractions from livers from experimental animals.

Studies on the effect of inflammation on rat serum proteins

1970 ◽  
Vol 48 (8) ◽  
pp. 841-850 ◽  
Author(s):  
F. E. Ashton ◽  
J. C. Jamieson ◽  
A. D. Friesen
Keyword(s):  
Serum Proteins ◽  
Deae Cellulose ◽  
Rat Serum ◽  
Experimental Animals ◽  
Inflammatory Agent ◽  
Fractionation Procedure ◽  
Time Periods ◽  
Short Time ◽  
New Protein ◽  
Short Times

The effect of turpentine-induced inflammation on rat serum proteins has been studied at short time periods of exposure to an inflammatory agent (5–96 h). There was an increase in protein-bound hexose and hexosamine of serum reaching a maximum at 48 h after administration of turpentine. Eight fractions were prepared from serum by a combination of chromatography on DEAE-cellulose and preparative electrophoresis on strips of gelatinized cellulose acetate. Most of the increase in protein-bound carbohydrate (77–86%) found in serum from experimental animals was located in three fractions following the fractionation procedure; immunological studies revealed the presence of fibrinogen and haptoglobin in two of these fractions. There was no evidence for the presence of a new protein in serum at short times of exposure to an inflammatory agent. Proteins present in the perchloric acid soluble and seromucoid fractions of serum were found in five fractions, only two of which contributed to the increase in protein-bound carbohydrate of serum found as a result of inflammation. Studies involving incorporation of labelled precursors of glycoprotein biosynthesis into fractions from serum indicated that the most likely explanation for the increase in protein-bound carbohydrate of serum, as a result of inflammation, was an increase in the rate of synthesis of certain serum glycoproteins.

scholarly journals Use of radioactive glucosamine in the perfused rat liver to prepare α1-acid glycoprotein (orosomucoid) with 3H- or 14C-labelled sialic acid and N-acetylglucosamine residues

1982 ◽  
Vol 203 (1) ◽  
pp. 141-148 ◽  
Author(s):  
N N Aronson
Keyword(s):  
Sialic Acid ◽  
Rat Liver ◽  
Specific Radioactivity ◽  
Deae Cellulose ◽  
Experimental System ◽  
Amino Sugar ◽  
Perfused Rat Liver ◽  
Rat Serum ◽  
Acid Glycoprotein ◽  
Sequential Digestion

1. A method was developed whereby [1-14C]glucosamine was used in a perfused rat liver system to prepare over 2 mg of alpha 1-acid glycoprotein with highly radioactive sialic acid and glucosamine residues. 2. The liver secreted radioactive alpha 1-acid glycoprotein over a 4-6 h period, and this glycoprotein was purified from the perfusate by chromatography on DEAE-cellulose at pH 3.6. 3. The sialic acid on the isolated glycoprotein had a specific radioactivity of 3.1 Ci/mol, whereas the glucosamine-specific radioactivity was 4.3 Ci/mole. The latter amino-sugar residues on the isolated protein were only 13-fold less radioactive than the initially added [1-14C]glucosamine. Orosomucoid with a specific radioactivity of 31.3 microCi/mg of protein was obtainable by using [6-3H]glucosamine. 4. The amino acid composition of the purified orosomucoid was comparable with that found by others for the same glycoprotein isolated from rat serum. A partial characterization of the carbohydrate structure was done by sequential digestion with neuraminidase, beta-D-galactosidase and beta-D-hexosaminidase. 5. Many other radioactive glycoproteins were found to be secreted into the perfusate by the liver. Thus this experimental system should prove useful for obtaining other serum glycoprotein with highly radioactive sugar moieties.

Studies on Acute Phase Proteins of Rat Serum. I. Isolation and Partial Characterization of an α1-Acid Glycoprotein and an α2-Macroglobulin

1972 ◽  
Vol 50 (8) ◽  
pp. 856-870 ◽  
Author(s):  
J. C. Jamieson ◽  
A. D. Friesen ◽  
F. E. Ashton ◽  
B. Chou
Keyword(s):  
Molecular Weight ◽  
Physical Properties ◽  
Acute Phase ◽  
Isoelectric Point ◽  
Acute Phase Proteins ◽  
Partial Characterization ◽  
Rat Serum ◽  
Acid Glycoprotein ◽  
Α2 Macroglobulin

An α1-acid glycoprotein and an α2-macroglobulin were isolated from serum from rats suffering from experimentally induced inflammation for 48 h. The proteins were characterized electrophoretically and immunologically and their carbohydrate compositions and some physical properties determined. The α1-acid glycoprotein contained 34.1% carbohydrate, had an isoelectric point of 2.95, and a molecular weight of 43 000. The α2-macroglobulin contained 15.9% carbohydrate, had an isoelectric point of 4.60, and a molecular weight of about 800 000. The properties of the two proteins are compared with those of similar proteins isolated from rats and other species.

scholarly journals Experimental Ehrlichia canis infection changes acute-phase proteins

2012 ◽  
Vol 21 (3) ◽  
pp. 206-212 ◽  
Author(s):  
Thiago Demarchi Munhoz ◽  
Joice Lara Maia Faria ◽  
Giovanni Vargas-Hérnandez ◽  
José Jurandir Fagliari ◽  
Áureo Evangelista Santana ◽  
...  
Keyword(s):  
Early Detection ◽  
Early Diagnosis ◽  
Acute Phase ◽  
Acute Phase Proteins ◽  
Ehrlichia Canis ◽  
Laboratory Findings ◽  
Early Detection Of Disease ◽  
Acid Glycoprotein ◽  
The Third ◽  
Canine Ehrlichiosis

Early diagnosis of canine ehrlichiosis favors prompt institution of treatment and improves the prognosis for the animal, since this disease causes mortality among dogs. Studies have shown that determining the concentration of acute-phase proteins (APPs) may contribute towards early detection of disease and aid in predicting the prognosis. This study aimed to evaluate the APP profile in dogs experimentally infected with Ehrlichia canis, at the start of the infection and after treatment. It also investigated whether any correlation between APP levels and the clinical and laboratory alterations over the course of the disease would be possible. The results obtained showed abnormal levels of all the APPs on the third day after infection (D3), with the highest levels being reached on D18, with the exception of ceruloplasmin and acid glycoprotein, which presented their peaks on D6 and D12 respectively. We concluded that assessment of APP levels could contribute towards establishing an early diagnosis of canine ehrlichiosis, particularly regarding acid glycoprotein and ceruloplasmin, since these proteins were detected at increased levels even before the onset of clinical and laboratory findings of the disease.

scholarly journals Luminometric assays of seven acute-phase proteins in minimal volumes of serum, plasma, sputum, and bronchioalveolar lavage.

1986 ◽  
Vol 32 (5) ◽  
pp. 743-747 ◽  
Author(s):  
J Braun ◽  
T Schultek ◽  
K F Tegtmeier ◽  
A Florenz ◽  
C Rohde ◽  
...  
Keyword(s):  
Acute Phase ◽  
Acute Phase Proteins ◽  
Immunoglobulin A ◽  
C Reactive Protein ◽  
Acid Glycoprotein ◽  
Reactive Protein ◽  
Coefficients Of Variation ◽  
Alpha 1 Antitrypsin ◽  
Heel Prick ◽  
Secretory Immunoglobulin

Abstract We describe immunoluminometric assays for seven acute-phase proteins, which can be determined in minimal volumes of plasma, serum, sputum, and bronchioalveolar lavage. The theoretical volume of serum or plasma required to measure all seven analytes in duplicate is 130 nL, although in practice the smallest volume of sample was enough to fill a hematocrit tube (about 25 microL of blood), collected from neonates by the heel-prick method. The assays could be performed with 10 microL of sputum or with 100 microL of bronchioalveolar lavage. We measured alpha 1-antitrypsin, alpha 2-macroglobulin, alpha 1-acid glycoprotein, thyroxin-binding prealbumin, C-reactive protein, and total and secretory immunoglobulin A. The assays are rapid enough for all results to be returned to the ward on the same day and are suitable for monitoring neonatal sepsis. All coefficients of variation, derived from compound precision profiles, were less than 7% for clinically relevant analyte concentrations. Correlation with commercially available nephelometric assays was good.

scholarly journals Effects of insulin in vitro on protein turnover in rat epitrochlearis muscle

1983 ◽  
Vol 210 (2) ◽  
pp. 323-330 ◽  
Author(s):  
W S Stirewalt ◽  
R B Low
Keyword(s):  
Protein Synthesis ◽  
Protein Degradation ◽  
Nitrogen Balance ◽  
Protein Metabolism ◽  
Serum Insulin ◽  
Specific Radioactivity ◽  
Physiological Concentration ◽  
Rat Serum ◽  
The Third

Rates of protein synthesis and degradation were measured in the isolated rat epitrochlearis muscle by radiotracer techniques, by using the specific radioactivity of tRNA-bound amino acid as precursor for protein synthesis. The tissue maintained linear rates of protein synthesis for 3 h of incubation in the presence of amino acids and glucose and in the absence of insulin. Under these conditions, however, the muscles were in negative nitrogen balance, with rates of protein degradation exceeding rates of protein synthesis. Under steady-state conditions of labelling, the specific radioactivities of tRNA-bound leucine, phenylalanine and valine were significantly less than their respective values in the incubation medium, at concentrations in the medium varying from 1 to 10 times those in normal rat serum. Insulin caused a dose- and time-dependent increase in tRNA-based protein synthesis rates, more than doubling rates at 5 and 50 ng of insulin/ml. At the lower, physiological, concentration of insulin, the stimulation of protein synthesis was not observed until the third hour of incubation with the hormone, whereas the rate of protein synthesis at the higher concentration was elevated during the second hour. There were no delays in the stimulation by insulin of glucose conversion into glycogen. The delayed stimulatory effects of insulin on the rate of protein synthesis brought the tissue to a nitrogen balance near zero. The presence of the hormone also prevented the increase in the rate of protein degradation seen in the third hour of incubation in the absence of the hormone. These studies demonstrate the viability of the incubated rat epitrochlearis muscle with respect to protein metabolism and sensitivity to the protein anabolic effects of physiological concentrations of insulin, and indicate that the preparation is a suitable experimental model for the study of the control of protein metabolism in fast-twitch skeletal muscle.

STUDIES ON THE SYNTHESIS OF PLASMA GLYCOLIPOPROTEIN AND HEPATIC SUB-CELLULAR GLYCOPROTEIN IN EARLY CHOLINE DEFICIENCY

1967 ◽  
Vol 45 (6) ◽  
pp. 825-838 ◽  
Author(s):  
Sailen Mookerjea ◽  
David Jeng ◽  
Judith Black
Keyword(s):  
Liver Microsome ◽  
Specific Radioactivity ◽  
Plasma Lipoproteins ◽  
Residual Fraction ◽  
Choline Deficiency ◽  
Glycoprotein Synthesis ◽  
Limiting Step ◽  
Residual Protein ◽  
Lipoprotein Synthesis ◽  
Smooth Membrane

Impairment of glycoprotein synthesis in trichloroacetic acid insoluble and in residual protein (d > 1.21) fractions of plasma in early choline deficiency has been confirmed. Plasma lipoproteins of d < 1.006, 1.006–1.063, and 1.063–1.21 are also rapidly labelled after intraperitoneal injection of glucosamine-1-14C. In general, there was no difference in specific radioactivity of plasma glycolipoproteins between choline-supplemented rats and rats deprived of choline for 2 days during 180 min of incorporation.Incorporation of glucosamine-1-14C into microsomal glycoprotein is significantly impaired in early choline deficiency. Further sub-microsomal fractionation produced evidence that the site of impairment of glycoprotein synthesis is in the smooth microsome portion. These studies suggest that a decreased synthesis of glycoprotein(s) in liver-microsome smooth membrane and in the plasma residual fraction (d > 1.21) may represent an impairment of 'lipoprotein precursor protein' synthesis in early choline deficiency. An inhibition of this limiting step would possibly impair the rate of plasma lipoprotein synthesis, leading to the development of fatty liver in choline deficiency.

Sign in / Sign up

Export Citation Format

Share Document