INHIBITION BY HUMAN SERUM OF THE ADIPOKINETIC EFFECT OF A HUMAN PITUITARY LIPID-MOBILIZING FACTOR

1972 ◽  
Vol 71 (3) ◽  
pp. 443-453 ◽  
Author(s):  
Olav Trygstad ◽  
Irene Foss
Keyword(s):  
Human Serum ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Inhibitory Effect ◽  
Disc Electrophoresis ◽  
Inhibitory Protein ◽  
Human Pituitary ◽  
Albumin Fraction ◽  
Pituitary Glands ◽  
Normal Human

ABSTRACT A lipid-mobilizing factor (LMF) with an adipotrophic effect in human and animal fat tissue has been prepared from human pituitary glands. The addition of normal human serum to LMF reduced its lipolytic effect, and it was completely abolished by serum from a group of obese patients, whereas the lipolysis was not influenced by serum from patients with generalized lipodystrophy. By DEAE-cellulose chromatography of human serum the inhibitory effect on LMF was found to be present in a protein fraction less acidic than the main serum albumin fraction. The inhibitory fraction was deprived of some contaminants by Sephadex gel filtration. Disc electrophoresis demonstrated the presence of three components in the inhibitory protein (IP), and they were identified as albumin, transferin, and haemopexin by immuno-electrophoresis. Precipitation of these proteins by their rabbit antisera demonstrated that the inhibitory effect was present in the albumin fraction. Insulin like activity was not observed in IP. A protein binding of LMF by IP could not be demonstrated. Incubation at 37°C for one hour of a mixture of LMF and IP eliminated the electrophoretic picture of LMF. It is concluded that the inhibitory effect of human serum may be due to proteolysis of LMF.

INHIBITION BY HUMAN SERUM OF THE ADIPOKINETIC EFFECT OF A HUMAN PITUITARY LIPID-MOBILIZING FACTOR (LMF) IN RABBITS

1967 ◽  
Vol 56 (4) ◽  
pp. 649-660 ◽  
Author(s):  
Olav Trygstad ◽  
Irene Foss
Keyword(s):  
Human Serum ◽  
Deae Cellulose ◽  
Globulin Fraction ◽  
Obese Patients ◽  
Human Pituitary ◽  
Nonesterified Fatty Acids ◽  
Pituitary Glands ◽  
Serum Inhibition ◽  
Normal Human ◽  
Metabolic Type

ABSTRACT A lipid-mobilizing factor (LMF) has previously been prepared from human pituitary glands. The LMF was strongly adipokinetic in rabbits. Subcutaneous injection of 0.1 mg gave an increase of serum nonesterified fatty acids (NEFA) in the range of 5 meq./l. Addition of 3 ml normal human serum to LMF reduced its adipokinetic effect. The inhibition of LMF by human serum was specific, since serum from mouse or pig did not reduce the increase of serum NEFA induced by LMF, and human serum had no influence on the adipokinesis given by whale, pig, or synthetic ACTH preparations in rabbits. The human serum inhibition of LMF was located to the gamma G globulin fraction by DEAE-cellulose chromatography. Serum from six children with generalized lipodystrophy had a negligible influence on the adipokinesis in rabbits induced by LMF. Sera from one group of obese patients reduced the adipokinetic effect of LMF as normal control sera, while sera from another group of obese patients made the increase of serum NEFA negligible. It is suggested that the first group of obese patients may be related to Mayer's simple regulatory or 'hyperphagic' type of obesity and the latter to his metabolic type of obesity. Furthermore possible mechanisms for the human serum inhibition of LMF are discussed.

ISOLATION OF LH, FSH AND TSH FROM HUMAN PITUITARIES AFTER THE REMOVAL OF HGH

1974 ◽  
Vol 77 (3) ◽  
pp. 485-497 ◽  
Author(s):  
P. A. Torjesen ◽  
T. Sand ◽  
N. Norman ◽  
O. Trygstad ◽  
I. Foss
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Disc Electrophoresis ◽  
Exchange Chromatography ◽  
Polyacrylamide Gel ◽  
Molecular Weights ◽  
Pituitary Glands

ABSTRACT Highly purified human LH, FSH and TSH were isolated from batches of 300 frozen pituitary glands (200 g) by pH, acetone and ethanol fractionation, Sephadex gel filtration, ion-exchange chromatography on DEAE-cellulose and CM-Sephadex, and preparative polyacrylamide-gel electrophoresis. Sodium dodecyl-sulphate (SDS) polyacrylamide gel electrophoresis was used in order to check the purity, the identity and the molecular weight of the purified LH, FSH and TSH. This procedure showed that the hormone preparations consisted of two subunits with molecular weights of: LH: 21 300 and 17 900, FSH: 22 100 and 18 300 and TSH: 20 800 and 16 400. The purity of the hormone preparations was also evaluated by analytical disc electrophoresis at pH 8.9. The purified hormone preparations had radioimmunological activity as follows: LH: 20 000 IU/mg, FSH: 16 500 IU/mg and TSH: 5 IU/mg. All preparations had high biological potency.

scholarly journals Purification and properties of human serum cholinesterase

1970 ◽  
Vol 116 (5) ◽  
pp. 875-881 ◽  
Author(s):  
P. K. Das ◽  
J. Liddell
Keyword(s):  
Human Serum ◽  
Catalytic Properties ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Disc Electrophoresis ◽  
Serum Cholinesterase ◽  
Purification And Properties ◽  
Preliminary Purification ◽  
Rare Genetic Variants ◽  
Human Cholinesterase

Cholinesterase was purified from human serum by a three-stage procedure involving chromatography on DEAE-cellulose at pH4.0, an electrofocusing technique and gel filtration on Sephadex G-200. The final product was purified 13000-fold with a yield of 54%, and only one protein and one cholinesterase band could be demonstrated by polyacrylamide-disc electrophoresis. The catalytic properties appeared to be unchanged by the purification procedure. The molecular weight was determined by both ultracentrifugation in a density gradient and gel filtration, and values close to 366000 were obtained. The isoelectric point of cholinesterase was estimated to be pH3.99. The method appears suitable for the preliminary purification of the rare genetic variants of human cholinesterase.

INHIBITION BY HUMAN SERUM OF THE ADIPOKINETIC EFFECT OF A HUMAN PITUITARY LIPID-MOBILIZING FACTOR

1973 ◽  
Vol 72 (2) ◽  
pp. 226-234 ◽  
Author(s):  
Olav Trygstad ◽  
Irene Foss
Keyword(s):  
Serum Albumin ◽  
Deae Cellulose ◽  
Disc Electrophoresis ◽  
Serum Albumins ◽  
Human Pituitary ◽  
Esterified Fatty Acids ◽  
Pituitary Glands ◽  
The Media ◽  
Fat Pads

ABSTRACT A lipid mobilizing factor (LMF) with an adipotrophic effect in man and animal adipose tissue has been prepared from human pituitary glands. In vitro studies have demonstrated that the lipolytic effect was dependent on the albumin used in the incubation medium. Two of the six albumins studied depressed the release of non-esterified fatty acids from human fat pads. Polyacrylamide gel disc-electrophoresis demonstrated the presence of contaminations in purchased serum albumins. They were eliminated by DEAE-cellulose chromatography, which gave one fraction that inhibited the adipokinetic effect of LMF, and a more homogeneous albumin fraction. In vitro lipolysis in rabbit as well as in human fat pads occurred more readily in the media containing purified serum albumin than in media with purchased serum albumin. It was concluded that in vitro lipolysis gives comparable results when studied in media containing the same batch of serum albumin only. Furthermore an agent should not be claimed to be non-lipolytic until it has been assayed in several, preferably homogeneous serum albumin media.

PURIFICATION AND PROPERTIES OF THE FOLLICLE-STIMULATING HORMONE INHIBITOR OBTAINED FROM THE URINE OF THE BONNET MONKEY

1968 ◽  
Vol 40 (2) ◽  
pp. 165-173 ◽  
Author(s):  
M. R. SAIRAM ◽  
H. G. MADHWA RAJ ◽  
N. R. MOUDGAL
Keyword(s):  
Follicle Stimulating Hormone ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Selective Extraction ◽  
Human Pituitary ◽  
Ammonium Sulphate Precipitation ◽  
Purification And Properties ◽  
Rat Pituitary ◽  
Pituitary Glands ◽  
Stimulating Hormone

SUMMARY The follicle-stimulating hormone (FSH) inhibitor in monkey urine was purified by selective extraction of the crude extract with acetate buffer, ammonium sulphate precipitation and DEAE-cellulose chromatography. The purified inhibitor was free of luteinizing hormone activity. It behaved as an apparently homogeneous protein. The inhibitor contained about 20% carbohydrate (hexoses, hexosamines, fucose and sialic acid). Thin-layer gel filtration indicated a molecular weight of about 65,000. The inhibitor was labile to heat treatment, exposure to extremes of pH and denaturing agents. The inhibitor effectively neutralized the biological activity of FSH preparations from human, monkey, horse, pig, sheep and rat pituitary glands, pregnant mare serum gonadotrophin and human pituitary urinary gonadotrophin.

Anticoagulants Produced by Thrombin from Fibrin, the Effect on Blood Coagulation, Some Physical Characteristics

1971 ◽  
Vol 26 (02) ◽  
pp. 211-223 ◽  
Author(s):  
Ch R. Muirhead ◽  
D. C Triantaphyllopoulos
Keyword(s):  
Blood Coagulation ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Fibrin Clot ◽  
Disc Electrophoresis ◽  
Physical Characteristics ◽  
Advanced Stage ◽  
Kallikrein Inhibitor ◽  
Shed Blood ◽  
Complete Lysis

SummaryChromatographed thrombin in the presence of both 50 Kallikrein inhibitor units of Trasylol per ml and 0.1 M E-ACA solubilized fibrin and the products of lysis possessed anticoagulant properties. The peak of the antithrombic activity coincided with the time of complete lysis of the fibrin clot, plasmin lysed fibrin exhibited the peak of its antithrombic activity much earlier. The effect of thrombin lysed fibrin on the prothrombin consumption of shed blood was found to be inhibitory.The products of the digestion of fibrin by thrombin and by plasmin, isolated at an advanced stage of proteolysis were compared by gel filtration, disc electrophoresis and DEAE cellulose chromatography. Differences in physical characteristics of these fibrin breakdown products offer evidence that they were produced by two different enzymes.

scholarly journals Purification and subunit structure of argininosuccinate lyase from Chlamydomonas reinhardi

1977 ◽  
Vol 167 (1) ◽  
pp. 71-75 ◽  
Author(s):  
R F Matagne ◽  
J P Schlösser
Keyword(s):  
Enzyme Activity ◽  
Crude Extract ◽  
Enzyme Preparation ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Activity Analysis ◽  
Disc Electrophoresis ◽  
Subunit Structure ◽  
Argininosuccinate Lyase

Argininosuccinate lyase (EC 4.3.2.1) was purified by (NH4)2SO4 fractionation, chromatography on DEAE-cellulose and gel filtration on Sephadex G-200. The final enzyme preparation was purified 46-fold compared with the crude extract. Electrophoresis of this preparation revealed three bands, the major one having the enzyme activity. Analysis of the enzyme by gel filtration and by disc electrophoresis (in two different concentrations of acrylamide) gave mol.wts. of 200000 (+/- 15000) and 190000 (+/- 20000) respectively. Treatment with sodium dodecyl sulphate and mercaptoethanol dissociated the enzyme into subunits of mol.wt. 39000 (+/-2000). The results are indicative of the multimeric structure of the enzyme, which is composed of five (perhaps four or six) identical subunits.

scholarly journals Purification and characterization of acid phosphatase from a germinating black gram (Vigna mungo L.) seedling

2011 ◽  
Vol 63 (3) ◽  
pp. 747-756 ◽  
Author(s):  
A.K.M. Asaduzzaman ◽  
Habibur Rahman ◽  
Tanzima Yeasmin
Keyword(s):  
Acid Phosphatase ◽  
Gel Electrophoresis ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Inhibitory Effect ◽  
Maximum Activity ◽  
Exchange Chromatography ◽  
Black Gram ◽  
Km Value

An acid phosphatase has been isolated and purified from an extract of a germinating black gram seedling. The method was accomplished by gel filtration of a germinating black gram seedling crude extract on sephadex G-75 followed by ion exchange chromatography on DEAE cellulose. The acid phosphatase gave a single band on SDS-polyacrylamide slab gel electrophoresis. The molecular weight of the acid phosphatase determined by SDS-polyacrylamide slab gel electrophoresis was estimated to be 25 kDa. The purified enzyme showed maximum activity at pH 5 and at temperature of 55?C. Mg2+, Zn2+ and EDTA had an inhibitory effect on the activity of the acid phosphatase. Black gram seedling acid phosphatase was activated by K+, Cu2+ and Ba2+. The Km value of the enzyme was found to be 0.49 mM for pNPP as substrate.

The Binding of Lithium Carmine to a2-Macroglobulin

1969 ◽  
Vol 24 (11) ◽  
pp. 1442-1447 ◽  
Author(s):  
J. J. Picard ◽  
J. F. Heremans
Keyword(s):  
Human Serum ◽  
Density Gradient ◽  
Protein Complex ◽  
Gel Filtration ◽  
Heavy Fraction ◽  
Normal Human Serum ◽  
Density Gradient Ultracentrifugation ◽  
Γ Globulins ◽  
Normal Human

The colloidal dye lithium carmine was added in vitro to normal human serum. Electrophoretic experiments showed that the dye was associated mainly with α2-globulins, small amounts with the albumin and only traces with the γ-globulins. The main complex was eluted with the macroglobulin peak obtained by gel filtration on Sephadex G-200 and sedimented in the heavy fraction on density gradient ultracentrifugation. The dye-protein complex could be precipitated with an antiserum specific for a2-macroglobulin. Gel filtration of a solution of pure a2-macroglobulin, to which lithium carmine was added, demonstrated that the dye was bound to this protein.

Sign in / Sign up

Export Citation Format

Share Document