scholarly journals The isolation of neurophysin-I and -II from bovine pituitary neurosecretory granules separated on a large scale from other subcellular organelles. Demonstration of slow equilibration of neurosecretory granules during centrifugation in a sucrose density gradient

1968 ◽  
Vol 106 (2) ◽  
pp. 565-573 ◽  
Author(s):  
C. R. Dean ◽  
D B Hope
Keyword(s):  
Pituitary Gland ◽  
Density Gradient ◽  
Biological Activities ◽  
Sucrose Density Gradient ◽  
Posterior Lobe ◽  
Starch Gel Electrophoresis ◽  
Neurosecretory Granules ◽  
Granule Protein ◽  
Pituitary Glands ◽  
Pressor Activity

1. An improved procedure for the isolation of neurosecretory granules from the posterior lobe of the bovine pituitary gland is described. 2. Of the total oxytocic and pressor activities present in the original tissue 80% was sedimentable. 3. The granules were separated from mitochondria by prolonged centrifugation in a sucrose density gradient. During a sedimentation period of 5hr. the granules moved progressively into denser regions of the gradient and the mitochondria remained at the top. 4. The biological activities of the granules were measured: the oxytocic activity was 11·56±1·63 and the pressor activity was 15·60±3·91 units/mg. of protein. 5. A protein was isolated from a lysate of granules prepared from 40 pituitary glands. Amino acid analysis showed that it consisted of a mixture of neurophysin-I and neurophysin-II in equal proportions. It accounted for 60% of the soluble granule protein and for 50% of the total granule protein. 6. The neurophysins present in the granules are associated with 19·1 units of oxytocic and 21·1 units of pressor activity/mg. of protein. 7. Starch-gel electrophoresis revealed the presence of both neurophysins in extracts of 15 pituitary glands studied individually. 8. We conclude that the polypeptide hormones, oxytocin and [8-arginine]-vasopressin, are normally closely associated with the two neurophysins within neurosecretory granules of the pituitary gland.

Crystallization of complexes of neurophysins with vasopressin and oxytocin

1968 ◽  
Vol 170 (1018) ◽  
pp. 37-47 ◽  
Keyword(s):  
Pituitary Gland ◽  
Biological Activities ◽  
Active Material ◽  
Posterior Lobe ◽  
Natural Form ◽  
Molecular Weights ◽  
Physico Chemical ◽  
Classical Work ◽  
Pressor Activity ◽  
Polypeptide Hormones

Shortly after the discovery of pressor activity in extracts of the posterior lobe of the pituitary gland (Oliver & Schafer 1895; Howell 1898) it was found that the addition of salt precipitated the active material (Osborne & Vincent 1900). Dale (1906) demonstrated the presence of the oxytocic principle in similar extracts. This activity was later found to accompany the pressor activity in salt precipitates (Kamm et al . 1928). Some years later van Dyke and his co-workers purified the material isolated from fresh pituitary tissue by salt precipitation. It was found to be a protein with a molecular weight of approximately 30 000, that appeared to be homogeneous by physico-chemical criteria. The protein possessed oxytocic, pressor and antidiuretic activities in the same ratios as they occur in the intact bovine pituitary gland (van Dyke, Chow, Greep & Rothen 1942). At that time, this protein was considered to be the natural form of a hormone possessing oxytocic, pressor and antidiuretic activities. However, another view, first expressed by Dudley (1919), suggested that the activities were attributable to the presence of two separate polypeptides. Kamm et al . (1928) extended the work of Dudley and by solvent extraction isolated two active principles, pitocin and pitressin, with molecular weights less than 1000. The classical work of du Vigneaud showed that the pituitary posterior lobe owes its characteristic biological activities to the presence of the polypeptide hormones, oxytocin and vasopressin (du Vigneaud 1955). The relation between the hormones and the so-called van Dyke protein was elucidated in Fromageot's laboratory. It was found that in mild acid oxytocin and vasopressin were dissociated from an inactive protein, neurophysin (Acher, Chauvet & Olivry 1956; Acher & Fromageot 1957. ) The portein-hormone complex could be reconstituted from the components in neutal solution.

DISTRIBUTION OF ADRENOCORTICOTROPHIC HORMONE IN THE PITUITARY GLAND

1957 ◽  
Vol 35 (1) ◽  
pp. 471-480
Author(s):  
G. J. Rochefort ◽  
M. Saffran
Keyword(s):  
Pituitary Gland ◽  
Relative Proportion ◽  
Posterior Lobe ◽  
Adrenocorticotrophic Hormone ◽  
Pituitary Glands

The highly vascular, anteromedial zone of hog and beef pituitary glands, which is rich in basophilic cells, contains a concentration of ACTH from 4 to 13 times that found in the rest of the adenohypophysis. The concentration of ACTH can be correlated (r = 0.96) with the relative proportion of basophilic cells, supporting the view that the pituitary basophiles are the source of ACTH. The distribution of both basophiles and ACTH in the rat adenohypophysis is more uniform than in hog or beef glands. The posterior lobe of all species examined (rat, hog, beef, and man) contains small, but significant, amounts of ACTH.

DISTRIBUTION OF ADRENOCORTICOTROPHIC HORMONE IN THE PITUITARY GLAND

1957 ◽  
Vol 35 (7) ◽  
pp. 471-480 ◽  
Author(s):  
G. J. Rochefort ◽  
M. Saffran
Keyword(s):  
Pituitary Gland ◽  
Relative Proportion ◽  
Posterior Lobe ◽  
Adrenocorticotrophic Hormone ◽  
Pituitary Glands

The highly vascular, anteromedial zone of hog and beef pituitary glands, which is rich in basophilic cells, contains a concentration of ACTH from 4 to 13 times that found in the rest of the adenohypophysis. The concentration of ACTH can be correlated (r = 0.96) with the relative proportion of basophilic cells, supporting the view that the pituitary basophiles are the source of ACTH. The distribution of both basophiles and ACTH in the rat adenohypophysis is more uniform than in hog or beef glands. The posterior lobe of all species examined (rat, hog, beef, and man) contains small, but significant, amounts of ACTH.

Isolation and characterization of β-lipolytic hormone from porcine pituitary gland

1970 ◽  
Vol 48 (9) ◽  
pp. 1017-1021 ◽  
Author(s):  
C. Gilardeau ◽  
M. Chrétien
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Pituitary Gland ◽  
Amino Acid Composition ◽  
Biological Activities ◽  
Terminal Amino Acid ◽  
Isolation And Characterization ◽  
Pituitary Glands ◽  
Terminal Amino

A lipolytic substance was isolated from porcine pituitary glands. It's amino acid composition, molecular weight, N-terminal amino acid, isoelectric point, and biological activities are reported. These results are compared to the corresponding values of sheep β-lipolytic hormone.

scholarly journals Solubilization of pig lymphocyte plasma membrane and fractionation of some of the components

1971 ◽  
Vol 123 (5) ◽  
pp. 967-975 ◽  
Author(s):  
D. Allan ◽  
M. J. Crumpton
Keyword(s):  
Plasma Membrane ◽  
Density Gradient ◽  
Density Gradient Centrifugation ◽  
Biological Activities ◽  
Gradient Centrifugation ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Sucrose Density Gradient ◽  
Similar Distribution ◽  
Sucrose Density Gradient Centrifugation

The degree of solubilization of pig lymphocyte plasma membrane by sodium deoxycholate was determined at a variety of temperatures and detergent concentrations. Approx. 95% of the membrane protein was soluble in 2% deoxycholate at 23°C. Some of the biological activities of the membrane survived this treatment. The leucine β-naphthylamidase activity was more readily soluble than the 5′-nucleotidase and these enzymes could be separated by extraction with 0.5% deoxycholate at 0°C. Membrane solubilized in 2% deoxycholate at 23°C was fractionated by sucrose-density-gradient centrifugation in 1% deoxycholate. The phospholipid was separated from the protein, which formed a fairly symmetrical peak that sedimented slightly slower than ovalbumin; the leucine naphthylamidase and 5′-nucleotidase activities were resolved from each other and from the main protein peak. Similar separations were achieved by elution from Sephadex G-200 and Sepharose 6B in 1% deoxycholate. The main proteins, however, appeared to possess much higher molecular weights than those indicated by sucrose-density-gradient centrifugation. This disparity suggests that many of the membrane proteins have a rod-like shape, especially since the results of experiments with [14C]deoxycholate revealed that the proteins did not bind significant amounts of deoxycholate. In contrast, 5′-nucleotidase and leucine naphthylamidase appeared to be globular. Polyacrylamide-gel electrophoresis of membrane solubilized in sodium dodecyl sulphate gave a similar distribution of protein to that achieved by sucrose-density-gradient centrifugation. Trace amounts only of polypeptides of molecular weight less than 10000 were detected.

THE MILK-EJECTING ACTIVITY OF EXTRACTS OF THE POSTERIOR PITUITARY GLAND

1952 ◽  
Vol 8 (2) ◽  
pp. 89-95 ◽  
Author(s):  
W. G. WHITTLESTONE
Keyword(s):  
Pituitary Gland ◽  
Posterior Lobe ◽  
Posterior Pituitary ◽  
High Potency ◽  
Pressor Activity ◽  
Posterior Pituitary Gland ◽  
Lactating Sow

The lactating sow milked mechanically has been used for the study of the hormones of the posterior pituitary gland. The van Dyke protein and du Vigneaud's high potency posterior lobe oxytocic polypeptide have been shown to possess milk-ejecting activity equivalent to their oxytocic activity. Commercial pressor preparations and du Vigneaud's highly purified vasopressin have milk-ejecting activity corresponding to a given pressor activity which is equivalent to one-fifth of the same number of oxytocic units.

The Combining Ability of Glycoproteins IIb, IIIa and Ca2+ in EDTA-Treated Nonaggregable Platelets

1983 ◽  
Vol 50 (04) ◽  
pp. 848-851 ◽  
Author(s):  
Marjorie B Zucker ◽  
David Varon ◽  
Nicholas C Masiello ◽  
Simon Karpatkin
Keyword(s):  
Density Gradient ◽  
Combining Ability ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Electrophoretic Pattern ◽  
Sucrose Density Gradient ◽  
Irreversible Loss ◽  
Sucrose Density Gradient Centrifugation ◽  
Crossed Immunoelectrophoresis ◽  
Triton X

SummaryPlatelets deprived of calcium and incubated at 37° C for 10 min lose their ability to bind fibrinogen or aggregate with ADP when adequate concentrations of calcium are restored. Since the calcium complex of glycoproteins (GP) IIb and IIIa is the presumed receptor for fibrinogen, it seemed appropriate to examine the behavior of these glycoproteins in incubated non-aggregable platelets. No differences were noted in the electrophoretic pattern of nonaggregable EDTA-treated and aggregable control CaEDTA-treated platelets when SDS gels of Triton X- 114 fractions were stained with silver. GP IIb and IIIa were extracted from either nonaggregable EDTA-treated platelets or aggregable control platelets with calcium-Tris-Triton buffer and subjected to sucrose density gradient centrifugation or crossed immunoelectrophoresis. With both types of platelets, these glycoproteins formed a complex in the presence of calcium. If the glycoproteins were extracted with EDTA-Tris-Triton buffer, or if Triton-solubilized platelet membranes were incubated with EGTA at 37° C for 30 min, GP IIb and IIIa were unable to form a complex in the presence of calcium. We conclude that inability of extracted GP IIb and IIIa to combine in the presence of calcium is not responsible for the irreversible loss of aggregability that occurs when whole platelets are incubated with EDTA at 37° C.

METABOLIC STUDIES WITH 131I-LABELED THYROXINE. II.

1963 ◽  
Vol 44 (3) ◽  
pp. 475-480 ◽  
Author(s):  
R. Grinberg
Keyword(s):  
Spinal Cord ◽  
Central Nervous System ◽  
Nervous System ◽  
Optic Nerve ◽  
Pituitary Gland ◽  
Time Interval ◽  
Time Intervals ◽  
Pituitary Glands ◽  
The Central Nervous System ◽  
Tentative Explanation

ABSTRACT Radiologically thyroidectomized female Swiss mice were injected intraperitoneally with 131I-labeled thyroxine (T4*), and were studied at time intervals of 30 minutes and 4, 28, 48 and 72 hours after injection, 10 mice for each time interval. The organs of the central nervous system and the pituitary glands were chromatographed, and likewise serum from the same animal. The chromatographic studies revealed a compound with the same mobility as 131I-labeled triiodothyronine in the organs of the CNS and in the pituitary gland, but this compound was not present in the serum. In most of the chromatographic studies, the peaks for I, T4 and T3 coincided with those for the standards. In several instances, however, such an exact coincidence was lacking. A tentative explanation for the presence of T3* in the pituitary gland following the injection of T4* is a deiodinating system in the pituitary gland or else the capacity of the pituitary gland to concentrate T3* formed in other organs. The presence of T3* is apparently a characteristic of most of the CNS (brain, midbrain, medulla and spinal cord); but in the case of the optic nerve, the compound is not present under the conditions of this study.

THE EXTRACTION OF PROLACTIN FROM HUMAN PITUITARY GLANDS

1965 ◽  
Vol 49 (1) ◽  
pp. 1-16 ◽  
Author(s):  
M. Apostolakis
Keyword(s):  
Growth Hormone ◽  
Pituitary Gland ◽  
List Type ◽  
Distilled Water ◽  
Hormone Activity ◽  
Isoelectric Precipitation ◽  
Human Pituitary ◽  
Pituitary Glands ◽  
Prolactin Content ◽  
The Mean

ABSTRACT A method for the extraction of prolactin from human pituitary glands is described. It is based on acetone drying, distilled water extraction, acetone and isoelectric precipitation. Two main products are obtained: Fraction R8 with a mean prolactin activity of 12.2 IU/mg and fraction U8 with a mean prolactin activity of 8.6 IU/mg. The former fraction does not contain any significant gonadotrophin activity and the latter contains on an average 50 HMG U/mg. In both cases contamination with ACTH and MSH is minimal. The growth hormone activity of both these fractions is low. It is postulated that in man too, prolactin and growth hormone are two distinct hormones. A total of 1250 human pituitary glands have been processed by this method. The mean prolactin content per pituitary gland has been found to be 73 IU.

FRACTIONATION AND ANALYSIS OF MONKEY PITUITARY GLANDS FOR POSTERIOR LOBE HORMONES

1962 ◽  
Vol 40 (2) ◽  
pp. 283-289 ◽  
Author(s):  
Darrell N. Ward ◽  
Earl F. Walborg ◽  
Harry S. Lipscomb ◽  
Roger Guillemin
Keyword(s):  
Amino Acid ◽  
Current Distribution ◽  
Arginine Vasopressin ◽  
Specific Activity ◽  
Small Scale ◽  
Posterior Lobe ◽  
Pharmacological Activities ◽  
Pituitary Glands ◽  
Counter Current ◽  
Counter Current Distribution

ABSTRACT Fractionation of monkey pituitary glands gave an oxytocin fraction in low yield which showed a counter-current distribution coefficient equivalent to that obtained with oxytocin from other species. Fractionation and chromatography of monkey vasopressin on carboxymethyl cellulose gave arginine-vasopressin of 60% purity, based on amino acid analysis and specific activity. Counter-current distribution on a small scale gave arginine-vasopressin of 89% purity. Reports by others that monkey pituitary glands contain arginine-vasopressin, based on pharmacological activities, are substantiated by the chemical data presented here.

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