Soluble esterases of human lung

1970 ◽  
Vol 48 (9) ◽  
pp. 970-978 ◽  
Author(s):  
J. Brebner ◽  
W. Kalow
Keyword(s):  
Molecular Weight ◽  
Column Chromatography ◽  
Human Lung ◽  
Human Tissues ◽  
Malic Dehydrogenase ◽  
Sephadex Column ◽  
Starch Gel

The soluble esterases of human lung compared to liver and serum have been illustrated on starch gel. A common zone of esterases appears to exist in nearly all human tissues and may be physiologically important acetylesterases. The most marked difference between lung and liver esterases exists in the ones moving farthest towards the anode at pH 7.0. Lactic and malic dehydrogenase patterns in the two tissues are also quite different. The zone 2 esterases appear to have a molecular weight of approximately 150 000 and those of zones 1 and 3 may be in the range of 50 000 as shown by G-200 Sephadex column chromatography.

STUDIES ON PLASMINOGEN: V. PURIFICATION OF BOVINE AND HUMAN PLASMINOGENS BY SEPHADEX CHROMATOGRAPHY

1966 ◽  
Vol 44 (4) ◽  
pp. 475-485 ◽  
Author(s):  
John Y. S. Chan ◽  
Edwin T. Mertz
Keyword(s):  
Human Serum ◽  
Column Chromatography ◽  
Continuous Flow ◽  
Sephadex Column ◽  
Molecular Weights ◽  
Starch Gel ◽  
Electrophoresis Method ◽  
Human Plasminogen ◽  
Specific Activities

Purified euglobulins prepared from bovine and human serum were fractionated by Sephadex column chromatography at pH 3.5. The products obtained were purified further by phosphate precipitation (bovine) or lysine solubilization (human) to yield final products with activities comparable to those prepared by the continuous-flow electrophoresis method. Plasminogen from Cohn fraction III-4 was also purified. The method yields preparations from serum which contain essentially no altered plasminogen. The specific activities, the starch-gel patterns, the molecular weights, and the recoveries of the purified bovine and human plasminogen preparations are reported and discussed.

EFFECT OF TRICHLORACETIC ACID ON BASOPHILS AND ON CORTICOTROPHIN CONTENT OF HUMAN PITUITARY GLANDS: EXTRACTION OF LOW MOLECULAR WEIGHT CORTICOTROPHIN

1963 ◽  
Vol 42 (1) ◽  
pp. 69-84 ◽  
Author(s):  
A. R. Currie ◽  
Beryl M. A. Davies
Keyword(s):  
Molecular Weight ◽  
Column Chromatography ◽  
Periodic Acid ◽  
Low Molecular Weight ◽  
Posterior Lobe ◽  
Human Pituitary ◽  
Trichloracetic Acid ◽  
Simple Method ◽  
Sephadex Column ◽  
Pituitary Glands

ABSTRACT Experiments are reported which show that 2.5% trichloracetic acid (TCA) causes marked loss of periodic acid-Schiff staining of the basophil granules of the pituitary gland and that there is extraction of some ACTH into the TCA. There is, however, no direct correlation between the basophil changes and the extraction of the hormone from the gland. The vesiculate chromophobe, the colloid and the cytoplasmic granules of the acidophils are not apparently affected by TCA treatment. Little is known about the chemical state of storage of ACTH in the human adenohypophysis. The amino acid structure and sequence of human ACTH has recently been proposed but there is as yet no chemical basis for a histochemical technique specific for ACTH. The 2.5% TCA-extractable ACTH, which represents about 30% of the total ACTH activity of batches of 'old' pituitary glands, has been characterized by ultrafiltration, Sephadex column chromatography and ultracentrifugation as a single component of minimum molecular weight about 3200. This is a simple method for extracting low molecular weight ACTH-extraction in 2.5% TCA (or water acidified to pH 5.0) for 1 hour at 4° C followed by ultrafiltration and Sephadex column chromatography. About 68% of the original activity in the TCA extract is recovered in the final product. Extraction was equally efficient whether 'pieces' or homogenates were used and in the presence or absence of posterior lobe tissue. Autolysis results in a decrease of both residual gland and TCA-extractable hormone.

PROLACTIN-LIKE ACTIVITY IN THE PITUITARY GLAND OF THE FROG

1966 ◽  
Vol 34 (2) ◽  
pp. 247-255 ◽  
Author(s):  
A. CHADWICK
Keyword(s):  
Pituitary Gland ◽  
Column Chromatography ◽  
Rana Temporaria ◽  
Sephadex Column ◽  
Zone Electrophoresis ◽  
Starch Gel ◽  
Distal Lobe

SUMMARY Homogenates of the distal lobe and combined intermediate and neural lobes from Rana temporaria and R. esculenta and extracts of these lobes, purified by zone electrophoresis in starch gel or by Sephadex column chromatography, were tested for crop-stimulating activity in the pigeon and lactogenic activity in the rabbit. The response of the pigeon crop to injections of homogenates resembled the reaction produced by purified sheep prolactin, but differed in intensity and in the number of fatty 'granules' present. The response in the rabbit differed considerably from the reaction to sheep prolactin. Milk was absent in spite of extensive proliferation in the gland. Injections of purified fractions of the pituitary extracts yielded similar results. It is concluded that a prolactin-like principle is present in the frog pituitary in both the distal lobe and the combined intermediate and neural lobes which can initiate a prolactin-like response in both the pigeon and the rabbit but cannot bring about the completion of a typical prolactin reaction.

scholarly journals Enzymic degradation of keratosulphates

1970 ◽  
Vol 119 (1) ◽  
pp. 39-47 ◽  
Author(s):  
Michiko Nishida-Fukuda ◽  
Fujio Egami
Keyword(s):  
Column Chromatography ◽  
Liver Extract ◽  
Concerted Action ◽  
Sephadex Column ◽  
Marine Gastropod ◽  
Multienzyme System ◽  
Shark Cartilage

1. A multienzyme system capable of degrading keratosulphates to yield galactose, N-acetylglucosamine and sulphate was found in the liver extract of a marine gastropod, Charonia lampas. 2. During the degradation, neither oligosaccharides nor sulphated sugars were produced. 3. It is suggested that the degradation could be attributed to the concerted action of β-galactosidase, β-N-acetylglucosaminidase and a sulphatase (sulphohydrolase), tentatively designated keratosulphatase. 4. Two forms of keratosulphatase (I and II) were separated by DEAE-Sephadex column chromatography. Both forms could release all the sulphate from keratosulphates and neither appeared to be identical with glycosulphatase or chondrosulphatase, both of which are also present in Charonia lampas. 5. β-Galactosidase and β-N-acetylglucosaminidase could degrade keratopolysulphate to a greater extent in the presence of keratosulphatase than in its absence. 6. It is suggested that keratosulphate was first desulphated by the action of keratosulphatase, and the desulphated polymer was then degraded to galactose and N-acetylglucosamine by the action of β-galactosidase and β-N-acetylglucosaminidase. 7. β-Galactosidase alone released a small amount of galactose from shark cartilage keratopolysulphate, but β-N-acetylglucosaminidase alone did not release N-acetylglucosamine. This indicates that unsulphated galactose residues occupy all the non-reducing terminal positions in keratopolysulphate chains.

scholarly journals A chromatographic preparation of ox growth hormone

1966 ◽  
Vol 100 (3) ◽  
pp. 593-600 ◽  
Author(s):  
M Wallis ◽  
HBF Dixon
Keyword(s):  
Molecular Weight ◽  
Growth Hormone ◽  
Gel Electrophoresis ◽  
Anterior Pituitary ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Starch Gel Electrophoresis ◽  
Highly Active ◽  
Cross Linkage ◽  
Starch Gel

1. A method is described for the chromatographic preparation of ox growth hormone. It involves chromatography of an extract of anterior pituitary lobes on DEAE-cellulose, followed by rechromatography on a dextran gel of low cross-linkage (Sephadex G-100). 2. The product is highly active in growth-hormone assays, and is obtained in good yield. It was homogeneous by several criteria, but showed some heterogeneity on starch-gel electrophoresis. 3. The molecular weight of the hormone was estimated from its behaviour on gel-filtration columns under various conditions. Evidence that the hormone may dissociate into sub-units under some conditions is presented.

Enzyme variation in the Anopheles gambiae Giles group of species (Diptera: Culicidae)

1978 ◽  
Vol 68 (1) ◽  
pp. 85-97 ◽  
Author(s):  
S. J. Miles
Keyword(s):  
Anopheles Gambiae ◽  
Natural Populations ◽  
Lactic Dehydrogenase ◽  
Starch Gel Electrophoresis ◽  
Malic Dehydrogenase ◽  
Glycerophosphate Dehydrogenase ◽  
Starch Gel ◽  
Anopheles Gambiae Giles ◽  
Species Specific ◽  
Glucose 6 Phosphate Dehydrogenase

AbstractThe genotypes of chromosomally-identified individuals from natural populations of the known species of the group of Anopheles gambiae Giles were scored for the enzyme protein structural loci coding for adenylate kinase (Adk), α-naphthyl acetate esterase (Est-1, Est-2, Est-3), glutamic-oxaloacetic transaminase (Got), α-glycerophosphate dehydrogenase (αGpd), hexokinase (Hk), isocitric dehydrogenase (Idh), lactic dehydrogenase (Ldh), ‘leucine’ aminopeptidase (Lap-2), malic enzyme (Me), octanol dehydrogenase (Odh), phosphoglucomutase (Pgm-1, Pgm-2), 6-phosphogluconic dehydrogenase (6-Pgd), phosphohexose isomerase (Phi) and superoxide dismutase (Sod), following starch gel electrophoresis. In the material examined, Est-1, Est-2, Est-3, Got, ldh, Lap-2, Odh, Pgm-1, Pgm-2 and Sod were segregating for two or more alleles; unique alleles at the Est-1, Got and Sod loci produced species-specific phenotypes in A. melas (Theo.), species C and species D, respectively. The further sampling of A. merus Dön, populations supported the presence of a unique SOD phenotype by which this species can also be identified. Of the other enzyme systems examined, no activity following electrophoresis was detected for aldolase and fructose-1,6-diphosphatase, and the resolution of acid and alkaline phosphatase, glyceraldehyde-3-phosphate dehydrogenase, glucose-6-phosphate dehydrogenase, malic dehydrogenase and xanthine dehydrogenase was too poor under the particular electrophoretic conditions for genetic analyses of the enzyme phenotypes.

scholarly journals Inhibition and complexation of activated protein C by two major inhibitors in plasma

Blood ◽  
1989 ◽  
Vol 73 (2) ◽  
pp. 446-454 ◽  
Author(s):  
MJ Heeb ◽  
F Espana ◽  
JH Griffin
Keyword(s):  
Molecular Weight ◽  
Column Chromatography ◽  
Protein C ◽  
Activated Protein C ◽  
Plasma Formation ◽  
The Other ◽  
Amidolytic Activity

Abstract To determine the major physiologic inhibitors of activated protein C (APC), plasma was incubated with APC or with Protac C and subjected to immunoblotting. APC:inhibitor complexes gave two major bands reacting with antiprotein C antibodies when immunoblotted on nondenaturing gels, and additional minor bands that varied between serum and plasma. Formation of one of the two major bands of APC:inhibitor complex, but not the other, was stimulated by heparin and only this band reacted with antibodies to the previously described APC inhibitor that is here designated PCI-1. Plasma immunodepleted of PCI-1 formed complexes with APC as visualized with antiprotein C but not anti-PCI-1 antibodies, and exhibited heparin-independent inhibition of APC activity, providing evidence for the existence of a second major physiologic APC inhibitor, PCI-2. Formation of APC:PCI-2 complexes in PCI-1-depleted plasma paralleled inhibition of APC amidolytic activity. PCI-2 was separated from PCI-1 and partially purified using column chromatography. PCI-2 formed inactive complexes of approximately 110,000 molecular weight (mol wt) with APC suggesting PCI-2 has an approximate mol wt of 50,000. Thus, inhibition of APC in plasma involves two major distinct 50,000 mol wt inhibitors, the heparin-dependent PCI-1 and the heparin- independent PCI-2.

scholarly journals Expression of TAL-1 proteins in human tissues

Blood ◽  
1995 ◽  
Vol 85 (3) ◽  
pp. 675-684 ◽  
Author(s):  
K Pulford ◽  
N Lecointe ◽  
K Leroy-Viard ◽  
M Jones ◽  
D Mathieu-Mahul ◽  
...  
Keyword(s):  
Molecular Weight ◽  
T Cell ◽  
Cell Lines ◽  
Fetal Liver ◽  
Cell Types ◽  
Human Tissues ◽  
Aberrant Expression ◽  
Erythroid Precursor ◽  
Normal Human

Rearrangement of the tal-1 gene (also known as SCL or TCL-5) occurs in at least 25% of T-cell acute lymphoblastic leukemias (T-ALLs) and results in the aberrant expression of tal-1 mRNA in the neoplastic cells. Also, tal-1 mRNA is constitutively expressed in erythroid precursors and megakaryocytes. This report describes a direct immunocytochemical study of the distribution and localization of TAL-1 protein in normal human tissues and cell lines using four monoclonal antibodies raised against recombinant TAL-1 proteins. One of these reagents recognizes a protein of 41 kD molecular weight in in vitro- translated TAL-1 proteins, two others recognize proteins of 39 and 41 kD molecular weight, and the fourth antibody also recognizes a TAL-1 protein of 22 kD in addition to the 39- and 41-kD proteins. These anti- TAL-1 antibodies label the nuclei of erythroid precursor cells and megakaryocytes in fetal liver and adult bone marrow. The punctate pattern of nuclear labeling suggests that TAL-1 may comprise part of a novel nuclear structure, similar to that recently found for the PML protein. The nuclei of T cell lines known to express mRNA encoding the full-length TAL-1 protein (eg, CCRF-CEM, RPMI 8402, and Jurkat) are also labeled. A study of normal human tissues (including thymus) showed labeling of smooth muscle, some tissue macrophages, and endothelial cells. TAL-1 protein is undetectable in other cell types. These reagents may play an important role in the diagnosis of T-ALL and could also be used in the context of lymphoma diagnosis on routinely fixed material.

scholarly journals Synthesis of adult-type hemoglobin in human erythremia cell line

Blood ◽  
1984 ◽  
Vol 64 (1) ◽  
pp. 314-317 ◽  
Author(s):  
M Kaku ◽  
K Yagawa ◽  
K Nakamura ◽  
H Okano
Keyword(s):  
Cell Line ◽  
Isoelectric Focusing ◽  
Column Chromatography ◽  
Cytosine Arabinoside ◽  
Adult Type ◽  
Sephadex Column ◽  
Cell Lysate ◽  
Globin Chains ◽  
Exposed Cell ◽  
Carboxy Methyl

Abstract KMOE -2/05 cells, derived from a patient with acute erythremia, became benzidine-positive after the addition of cytosine arabinoside (CA). Radioimmunoassays using antihuman hemoglobin antibodies revealed an elevated amount of hemoglobin in the CA-exposed cells over that in the control cells (without CA). Isoelectric focusing of the CA-exposed cell lysate formed benzidine-positive foci in the positions of human adult Hb (HbA) and human fetal Hb (HbF). To determine the types of globin synthesized in the CA-exposed cells, globin chains internally labeled with [3H] leucine were purified by carboxy-methyl (CM)-Sephadex column chromatography, immunoadsorption by Sepharose-coupled antihuman Hb antibodies and Sephadex G-100. The labeled globin chains were finally separated by CM-cellulose chromatography in urea. Two distinct peaks of radioactivity were shown in exactly the same fractions as carrier human globin alpha- and beta-chains. These observations indicate that these KMOE -2/05 cells synthesize HbA.

PROPERTIES AND CLASSIFICATION OF THE SOLUBLE ESTERASES OF HUMAN BRAIN

1966 ◽  
Vol 44 (2) ◽  
pp. 225-232 ◽  
Author(s):  
D. J. Ecobichon
Keyword(s):  
Alkaline Phosphatase ◽  
Human Brain ◽  
Esterase Activity ◽  
Water Soluble ◽  
Human Tissues ◽  
Esterase Pattern ◽  
Vertical Zone ◽  
Zone Electrophoresis ◽  
Starch Gel

Water-soluble proteins and enzymes of human brain were separated by vertical zone electrophoresis in starch gel. Fifteen bands of esterase activity were detected in brain. Various substrates and inhibitors were used in efforts to identify enzymes in addition to a comparison of the esterase pattern with patterns obtained from other human tissues. One zone, composed of four bands of acetylesterase activity, was found to be common to all the tissues investigated with the exception of serum. Two bands of cholinesterase and two bands of A-esterase activity were identified. The remaining bands, which were aliesterases possessing broad overlapping substrate specificity and inhibitor sensitivity, were electrophoretically different from those of other tissues. Observations on alkaline phosphatase, acid phosphatase, and lactate dehydrogenase were recorded for comparison with the data on esterases.

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