THYROID-STIMULATING AND LIPOLYTIC ACTIVITIES OF PURIFIED PREPARATIONS OF HUMAN THYROID-STIMULATING HORMONE

1972 ◽  
Vol 53 (1) ◽  
pp. 95-100 ◽  
Author(s):  
ANNE STOCKELL HARTREE ◽  
MARJORIE THOMAS ◽  
BRIDGET E. FURNIVAL ◽  
T. W. BURNS ◽  
P. LANGLEY
Keyword(s):  
Lipolytic Activity ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Protein Molecule ◽  
Thyroid Stimulating Hormone ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Human Thyroid ◽  
Rat Adipose Tissue ◽  
Stimulating Hormone

SUMMARY A partially purified fraction of human thyroid-stimulating hormone (DEAE-II) was further purified by ion-exchange chromatography on IRC-50, gel-filtration on Sephadex G-100 and finally chromatography on DEAE-cellulose. Two fractions were obtained which were high in thyroid-stimulating activity (8·3 and 7·3 units human Research Standard A/mg) and were comparable in potency to other preparations of the human hormone reported in the literature. They were also electrophoretically heterogeneous as were the preparations of other workers. Lipolytic activity toward cells obtained from human or rat adipose tissue was demonstrated for all fractions containing thyroid-stimulating activity, the two activities being roughly parallel. It is concluded that both thyroid-stimulating and lipolytic activities are probably present in the same protein molecule, but it is unlikely that the latter activity is of physiological significance.

scholarly journals The isocitrate dehydrogenases of Acinetobacter lwoffi. Separation and properties of two nicotinamide–adenine dinucleotide phosphate-linked isoenzymes

1972 ◽  
Vol 130 (1) ◽  
pp. 211-219 ◽  
Author(s):  
Colin H. Self ◽  
P. David J. Weitzman
Keyword(s):  
Molecular Weight ◽  
Ph Dependence ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
Molecular Size ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Low Concentrations ◽  
Stimulation Of

Two isoenzymes of NADP-linked isocitrate dehydrogenase have been identified in Acinetobacter lwoffi and have been termed isoenzyme-I and isoenzyme-II. The isoenzymes may be separated by ion-exchange chromatography on DEAE-cellulose, by gel filtration on Sephadex G-200, or by zonal ultracentrifugation in a sucrose gradient. Low concentrations of glyoxylate or pyruvate effect considerable stimulation of the activity of isoenzyme-II. The isoenzymes also differ in pH-dependence of activity, kinetic parameters, stability to heat or urea and molecular size. Whereas isoenzyme-I resembles the NADP-linked isocitrate dehydrogenases from other organisms in having a molecular weight under 100000, isoenzyme-II is a much larger enzyme (molecular weight around 300000) resembling the NAD-linked isocitrate dehydrogenases of higher organisms.

Purification of glysojanin, an antifungal protein, from the black soybean Glycine soja

2003 ◽  
Vol 81 (6) ◽  
pp. 387-394 ◽  
Author(s):  
Patrick H.K Ngai ◽  
T B Ng
Keyword(s):  
Liquid Chromatography ◽  
Ion Exchange ◽  
Glycine Soja ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Fast Protein Liquid Chromatography ◽  
Antifungal Protein ◽  
Black Soybean

A monomeric protein, with a molecular mass of 25 kDa and an N-terminal sequence resembling a segment of chitin synthase, was isolated from the seeds of the black soybean Glycine soja. The protein, designated glysojanin, demonstrated potent antifungal activity against the fungi Fusarium oxysporum and Mycosphaerella arachidicola. It inhibited HIV-1 reverse transcriptase with an IC50 of 47 µmol/L, [methyl-3H]thymidine incorporation by mouse spleen cells with an IC50 of 175 µmol/L, and translation in the rabbit reticulocyte lysate with an IC50 of 20 µmol/L. Glysojanin was purified using a procedure that involved ion-exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, ion-exchange chromatography by fast protein liquid chromatography on Mono S, and gel filtration by fast protein liquid chromatography on Superdex 75.Key words: antifungal protein, seeds, soybean, purification.

Comparative studies of manganese(II)-, nickel(II)-, zinc(II)-, copper(II)-, cadmium(II)-, and iron(III)-binding components in human cord and adult sera

1984 ◽  
Vol 62 (6) ◽  
pp. 449-455 ◽  
Author(s):  
Show-Jy Lau ◽  
Bibudhendra Sarkar
Keyword(s):  
Molecular Weight ◽  
Trace Metals ◽  
Ion Exchange ◽  
Comparative Studies ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Low Molecular Weight ◽  
High Molecular Weight Proteins

The binding of six trace metals, Mn(II), Ni(II), Zn(II), Cu(II), Cd(II) and Fe(III), to human cord serum has been studied by Sephadex G-100 gel filtration at physiological pH, using radioisotopes as tracers. The results are compared with those obtained from adult serum. In both cord and adult sera, extensive amounts of the metals are bound to high molecular weight proteins. Among them, Fe(III) is mostly bound to transferrin; Ni(II), Zn(II), Cu(II), and Cd(II) are bound to albumin and other macro-molecules. The binding of Mn(II) either to transferrin or albumin is not resolved. Small fractions of Zn(II), Cu(II), and Cd(II) and large fractions of Mn(II) and Ni(II) are found to be associated with low molecular weight components of both sera. The distribution varies from metal to metal. However, the low molecular weight component of the size 1500 – 10 000 is present in all the metals studied. Further purification of this component was attempted by DEAE-cellulose ion-exchange chromatography. The possible identity as well as the biological role played by this particular component of serum in the transport of metals in blood and across membranes is discussed.

Isolation of deoxyribonuclease II from bovine intestinal mucosa

1980 ◽  
Vol 58 (9) ◽  
pp. 749-753 ◽  
Author(s):  
D. Stephen Keys ◽  
S. H. Zbarsky
Keyword(s):  
Gel Filtration ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Degraded Dna ◽  
Major Peak ◽  
Dnase Ii ◽  
Minor Peak ◽  
Supernatant Solution ◽  
Bovine Small Intestine

Mucosa from bovine small intestine was homogenized in Krebs–Ringer phosphate buffer, pH 7.8, the homogenate centrifuged at 16 300 × g, and the supernatant solution filtered through cheesecloth to remove lipid material. The filtrate was centrifuged at 105 000 × g and the supernatant solution chromatographed on DEAE-cellulose. The major peak of DNase II activity, eluted with 20 mM phosphate – 10 mM EDTA buffer, pH 7.8, was purified further by ion-exchange chromatography on CM-cellulose and gel filtration on Sephadex G-100. The enzyme was purified 78-fold in 13% yield. Evidence was adduced to indicate that the second minor peak of DNase II activity, eluted from the DEAE-cellulose by a potassium chloride gradient in the 20 mM phosphate – 10 mM EDTA buffer, was an artifact arising from the presence of significant amounts of DNA in the 105 000 × g supernatant. The enzyme degraded DNA endonucleolytically to 3′-PO4, 5′-OH oligonucleotides and is similar in its properties to DNase II from other tissues.

Studies on salivary and pancreatic lipases of the pre-ruminant calf

1982 ◽  
Vol 49 (3) ◽  
pp. 347-360 ◽  
Author(s):  
Joyce Toothill
Keyword(s):  
Ion Exchange ◽  
Affinity Chromatography ◽  
Pancreatic Lipase ◽  
Lipase Activity ◽  
Lipolytic Activity ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Maximum Activity ◽  
Exchange Chromatography ◽  
Single Enzyme

SUMMARYSalivary and pancreatic lipases of the pre-ruminant calf have been studied using ion-exchange chromatography and gel filtration. In addition, pancreatic lipase has been fractionated using concanavalin A-affinity chromatography. The effects of 5,5′-dithiobis(2-nitrobenzoic acid), organic solvents and trypsin on pancreatic lipase have also been investigated. The effect of taurodeoxycholate on the lipolytic activity of the 2 lipases has been compared. Salivary lipase behaved as a single enzyme on ion-exchange chromatography, and gel filtration gave a mol. wt value of 52000 for the enzyme. Although pancreatic lipase appeared to be a single enzyme on gel filtration, with a mol. wt of almost 80000, the lipase was shown by ion-exchange and affinity chromatography to consist of at least 2 enzymes of mol. wts 72000 and 60000, and did not require colipase for maximum activity in the presence of high concentrations of bile salts. Colipase-dependent lipase, mol. wt about 45000, and probably amounting to not more than 10% of the total activity, was also present. This was the predominant form only after large losses in total lipolytic activity had occurred, as after treatment with a mixture of ether, ethanol and deoxycholate, or prolonged action of trypsin. When the concentration of taurodeoxycholate was increased from zero to 1 mM in a tributyrin substrate, the lipolytic activites of calf salivary and pancreatic lipases, and pig pancreatic lipase, increased. At a concentration of 4 mM-taurodeoxycholate, calf salivary lipase activity was higher, that of calf pancreatic lipase slightly lower and pig pancreatic lipase activity markedly lower.

scholarly journals Biochemical properties and positional specificity of Lipase from hepatopancreas of Tra catfish (Pangasius)

2013 ◽  
Vol 16 (4) ◽  
pp. 85-91
Author(s):  
Thy Bao Vuong ◽  
Lam Bich Tran ◽  
Duan Luu
Keyword(s):  
Ion Exchange ◽  
Lipase Activity ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Biochemical Properties ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Positional Specificity ◽  
Tra Catfish ◽  
Ph Range

Lipase from the hepatopancreas of Tra catfish (Pangasius) was precipitated by ammonium sulfate fractionation, purified by ion-exchange chromatography on DEAE cellulose and gel filtration on Sephadex G- 75. On substrate triolein, purified lipase has Km= 1.381 mM and Vmax= 0.063 mM/min. The lipase was stable at a pH range of 7.0- 9.0 and in temperatures of 35-50°C. At 500C the enzyme loosed 44,7% activity after 120 min. The enzyme was specific for the α- positions (1, 3) of triglyceride. In bile salt solution of 0.015M NaTC, lipase activity of the enzyme increased in 3.08 folds in comparison of sample without NaTC.

scholarly journals Novel galactonic acid-binding hexameric lectin from Hibiscus mutabilis seeds with antiproliferative and potent HIV-1 reverse transcriptase inhibitory activities.

2009 ◽  
Vol 56 (4) ◽  
Author(s):  
Sze Kwan Lam ◽  
Tzi Bun Ng
Keyword(s):  
Reverse Transcriptase ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Inhibitory Activities ◽  
Binding Lectin ◽  
Hiv 1 ◽  
Hepatoma Hepg2 ◽  
Mcf 7

A hexameric 150-kDa lectin was isolated from dried Hibiscus mutabilis seeds using a chromatographic protocol that involved ion exchange chromatography on SP-Sepharose, and gel filtration on Superdex 75 and Superdex 200. The lectin was not adsorbed on SP-Sepharose and was eluted from the Superdex 75 column in the void volume. It was eluted in the first peak from Superdex 200. It was strongly adsorbed on DEAE-cellulose and Q-Sepharose and could not be easily desorbed. The hemagglutinating activity of the lectin, which was stable at pH 4-7 and up to 50 degrees C, could be inhibited by 25 mM galactonic acid. This is the first report of a galactonic acid-binding lectin. It potently inhibited HIV-1 reverse transcriptase with an IC(50) of 0.2 microM. It exhibited weak antiproliferative activity towards both hepatoma HepG2 cells (40% inhibition) and breast cancer MCF-7 cells (50% inhibition) at 100 microM concentration of the lectin. It did not inhibit mycelial growth of a number of fungi tested.

scholarly journals Purification and amino acid sequence of melanocyte-stimulating hormone from the dogfish Squalus acanthias

1970 ◽  
Vol 118 (5) ◽  
pp. 713-718 ◽  
Author(s):  
P. J. Lowry ◽  
A. Chadwick
Keyword(s):  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Squalus Acanthias ◽  
Tyrosine Residue ◽  
Melanocyte Stimulating Hormone ◽  
C Terminus ◽  
N Terminus ◽  
Pituitary Glands ◽  
Stimulating Hormone

A melanocyte-stimulating hormone (MSH) was isolated by gel filtration and ion-exchange chromatography from extracts of the pituitary glands of dogfish. Sequence studies were carried out on the hormone and its enzymically and chemically cleaved fragments. The sequence of the hormone, Ser-Met-Glu-His-Phe-Arg-Trp-Gly-Lys-Pro-Met, shows that ten of its 11 residues are the same as ten of the 13 residues of mammalian α-MSH. About half of its molecules have the carboxyl group at the C-terminus free and about half are amidated; about a fifth have an extra tyrosine residue on the N-terminus, thereby making 11 residues the same as in mammalian α-MSH. Unlike the mammalian hormone, however, none of it was found to be N-acetylated.

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.

scholarly journals Trypsin Isoinhibitors with Antiproliferative Activity toward Leukemia Cells fromPhaseolus vulgariscv “White Cloud Bean”

2010 ◽  
Vol 2010 ◽  
pp. 1-8 ◽  
Author(s):  
Jian Sun ◽  
Hexiang Wang ◽  
Tzi Bun Ng
Keyword(s):  
Ion Exchange ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Fungal Growth ◽  
Inhibitory Effect ◽  
Ion Exchange Chromatography ◽  
Trypsin Inhibitors ◽  
Exchange Chromatography ◽  
L1210 Cells ◽  
Trypsin Inhibitory Activity

A purification protocol that comprised ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, ion exchange chromatography on SP-Sepharose, and gel filtration by FPLC on Superdex 75 was complied to isolate two trypsin inhibitors fromPhaseolus vulgariscv “White Cloud Bean”. Both trypsin inhibitors exhibited a molecular mass of 16 kDa and reduced the activity of trypsin with an value of about 0.6 M. Dithiothreitol attenuated the trypsin inhibitory activity, signifying that an intact disulfide bond is indispensable to the activity. [Methyl-] thymidine incorporation by leukemia L1210 cells was inhibited with an value of 28.8 M and 21.5 M, respectively. They were lacking in activity toward lymphoma MBL2 cells and inhibitory effect on HIV-1 reverse transcriptase and fungal growth when tested up to 100 M.

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