ISOLATION AND CHARACTERIZATION OF INHIBIN FROM BULL SEMINAL PLASMA

1978 ◽  
Vol 87 (2) ◽  
pp. 434-448 ◽  
Author(s):  
S. Chari ◽  
S. Duraiswami ◽  
P. Franchimont
Keyword(s):  
Seminal Plasma ◽  
Gel Filtration ◽  
Male Rats ◽  
Acetone Powder ◽  
Alkaline Ph ◽  
Sodium Acetate Buffer ◽  
Immature Rat ◽  
Immature Male ◽  
Isolation And Characterization

ABSTRACT A simple, short procedure for the isolation of inhibin from bull seminal plasma has been described. Following conversion of the seminal plasma into acetone powder, it was subjected to gel filtration on Sephadex G-100 using 4 m urea—0.05 m sodium acetate buffer pH 4.0 as the eluent. Further purification of the active fraction was achieved by 'straight elution analysis' using CM-cellulose. The final preparation (Ux) has been shown to reduce the response of immature rat ovaries to human chorionic gonadotrophic hormone (HCG) stimulation in the bioassay system of Chari et al. (1976) and also been shown to suppress the immediate post-castration rise in serum gonadotrophin levels in immature male rats. Homogeneity of the isolated fraction has been assessed electrophoretically at different pH's above and below its isoelectric point. On the basis of gel filtration studies, electrophoresis and amino acid analysis, the molecular weight of Ux has been estimated to be 19 000 daltons. The electrophoretic behaviour of Ux at an alkaline pH has been discussed as a possible evidence for its existence as isohormones.

Isolation and Characterization of Plasminogen Activator from Pig Leucocytes

1974 ◽  
Vol 31 (01) ◽  
pp. 072-085 ◽  
Author(s):  
M Kopitar ◽  
M Stegnar ◽  
B Accetto ◽  
D Lebez
Keyword(s):  
Molecular Weight ◽  
Plasminogen Activator ◽  
Gel Electrophoresis ◽  
Gel Filtration ◽  
Ph Optimum ◽  
Isolation And Characterization ◽  
Ph Range ◽  
Inhibition Studies ◽  
Final Purification

SummaryPlasminogen activator was isolated from disrupted pig leucocytes by the aid of DEAE chromatography, gel filtration on Sephadex G-100 and final purification on CM cellulose, or by preparative gel electrophoresis.Isolated plasminogen activator corresponds No. 3 band of the starting sample of leucocyte cells (that is composed from 10 gel electrophoretic bands).pH optimum was found to be in pH range 8.0–8.5 and the highest pH stability is between pH range 5.0–8.0.Inhibition studies of isolated plasminogen activator were performed with EACA, AMCHA, PAMBA and Trasylol, using Anson and Astrup method. By Astrup method 100% inhibition was found with EACA and Trasylol and 30% with AMCHA. PAMBA gave 60% inhibition already at concentration 10–3 M/ml. Molecular weight of plasminogen activator was determined by gel filtration on Sephadex G-100. The value obtained from 4 different samples was found to be 28000–30500.

scholarly journals α-Crystallin. The isolation and characterization of distinct macromolecular fractions

1971 ◽  
Vol 124 (2) ◽  
pp. 337-343 ◽  
Author(s):  
Abraham Spector ◽  
Lu-Ku Li ◽  
Robert C. Augusteyn ◽  
Arthur Schneider ◽  
Thomas Freund
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Chemical Difference ◽  
Molecular Weights ◽  
Isolation And Characterization ◽  
Different Populations ◽  
Molecular Weight Fractions

α-Crystallin was isolated from calf lens periphery by chromatography on DEAE-cellulose and gel filtration. Three distinct populations of macromolecules have been isolated with molecular weights in the ranges approx. 6×105−9×105, 0.9×106−4×106and greater than 10×106. The concentration of macromolecules at the molecular-weight limits of a population are very low. The members of the different populations do not appear to be in equilibrium with each other. Further, in those molecular-weight fractions investigated, no equilibrium between members of the same population was observed. The population of lowest molecular weight comprises 65–75% of the total material. The amino acid and subunit composition of the different-sized fractions appear very similar, if not identical. The only chemical difference observed between the fractions is the presence of significant amounts of sugar in the higher-molecular-weight fractions. Subunit molecular weights of approx. 19.5×103and 22.5×103were observed for all α-crystallin fractions.

PURIFICATION OF "INHIBIN" FROM HUMAN OVARIAN FOLLICULAR FLUID

1979 ◽  
Vol 90 (1) ◽  
pp. 157-166 ◽  
Author(s):  
S. Chari ◽  
C. R. N. Hopkinson ◽  
E. Daume ◽  
G. Sturm
Keyword(s):  
Follicular Fluid ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Exchange Chromatography ◽  
Male Rats ◽  
Acetone Powder ◽  
Gel Chromatography ◽  
Apparent Homogeneity ◽  
Ovarian Follicular Fluid

ABSTRACT Following the earlier demonstration of inhibin-like activity in human ovarian follicular fluid a method for its purification to apparent homogeneity is described. The fluid was converted to acetone powder and subjected sequentially to ammonium sulphate fractionation, gel chromatography on Sephadex G-200, continuous gradient ion-exchange chromatography on DEAE-cellulose, first with a pH gradient from 8.0 to 4.0 and then with a NaCl gradient to 1 m at pH 5.2. The active fraction from this step was subjected to gel filtration on Sephadex G-100 and finally passed through an Amicon Centriflo membrane CF-25 (cut off point: 25 000 m.w.). The ultrafiltrate was homogeneous by SDS-polyacrylamide gel electrophoresis, had a molecular weight of the order of 23 000 and was capable of suppressing serum gonadotrophin levels in the castrated male rats in as low a dose as 25 μg/rat.

scholarly journals Isolation and characterization of a novel haemoprotein b-559 from Bengal gram (Cicer arietinum)

1987 ◽  
Vol 243 (3) ◽  
pp. 723-728 ◽  
Author(s):  
C S Ramadoss ◽  
B C Shenoy ◽  
A Borthakur
Keyword(s):  
Gel Filtration ◽  
Molecular Size ◽  
Soret Band ◽  
Beta Band ◽  
Photosynthetic Membrane ◽  
Isolation And Characterization ◽  
Bengal Gram ◽  
Apparent Homogeneity ◽  
Reduced State

A haemoprotein was purified to apparent homogeneity from Bengal-gram seeds. The purified protein exhibited an absorption maximum at 412 nm (Soret band) that upon reduction with dithionite gave rise to a shift in the Soret band to 426 nm with concomitant appearance of an alpha-band at 559 nm and a beta-band at 530 nm. In the reduced state the Bengal-gram haemoprotein showed reactivity towards CO, nitrite and hydroxylamine. SDS/polyacrylamide-slab-gel electrophoresis showed that the haemoprotein has Mr 78,000. Gel-filtration and ultracentrifugal analyses suggest that the Bengal-gram haemoprotein is oligomeric in nature. Since it differs from photosynthetic membrane cytochrome b-559 in solubility in buffer, in reactivity towards CO and in molecular size, it appears to be a novel haemoprotein b-559.

Isolation and characterization of the major proteins of ram seminal plasma

2005 ◽  
Vol 71 (4) ◽  
pp. 461-470 ◽  
Author(s):  
Annick Bergeron ◽  
Michèle Villemure ◽  
Claude Lazure ◽  
Puttaswamy Manjunath
Keyword(s):  
Seminal Plasma ◽  
Isolation And Characterization

scholarly journals Novel heparin degradation products. Isolation and characterization of novel disaccharides and oligosaccharides produced from heparin by bacterial degradation

1968 ◽  
Vol 108 (4) ◽  
pp. 647-654 ◽  
Author(s):  
Carl P. Dietrich
Keyword(s):  
Paper Chromatography ◽  
Uronic Acid ◽  
Degradation Products ◽  
Gel Filtration ◽  
Present Knowledge ◽  
Bacterial Degradation ◽  
Isolation And Characterization ◽  
Flavobacterium Heparinum

1. Heparin was degraded by enzymes of adapted Flavobacterium heparinum. Several degradation products were separated by combined Sephadex-gel filtration and paper chromatography, and chemically analysed. 2. These products were identified as glucosamine 2,6-disulphate, saturated disaccharides constituted of uronic acid and glucosamine and containing two and three sulphate residues, and tetra- and hexa-saccharides with the same basic disaccharide units. 3. The implications of these findings with respect to the present knowledge of heparin structure and its enzymic degradation are discussed.

scholarly journals Isolation and characterization of membrane proteins responsible for attachment of polyribosomes to rough microsomal fraction of rat liver

1977 ◽  
Vol 164 (1) ◽  
pp. 53-66 ◽  
Author(s):  
S Fujita ◽  
F Ogata ◽  
J Nakamura ◽  
S Omata ◽  
H Sugano
Keyword(s):  
Rat Liver ◽  
Protein Fraction ◽  
Microsomal Fraction ◽  
Binding Capacity ◽  
Gel Filtration ◽  
High Affinity ◽  
Isolation And Characterization ◽  
Microsomal Membranes ◽  
Equilibrium Centrifugation

A protein fraction which has a high affinity for polyribosomes was isolated from rough microsomal membranes of rat liver. The mode of polyribosome binding to this fraction (R-fraction) was studied by using CsCl equilibrium centrifugation and compared with that for stripped rough microsomal membranes. The following were found. (1) The polyribosome-binding cpacity of the R-fraction was heat-labile and sensitive to trypsin, and was suppressed by increasing KCl concentration and addition of 0.1 mM-aurintricarboxylic acid. (2) Of the four subfractions obtained by gel filtration of the R-fraction on a Sephadex G-200, only the R1-fraction, eluted at the void volume, showed a high affinity for polyribosomes. The polyribosome-binding capacity of the R1-fraction decreased with time on storage at 4 degrees C. (3) The R1-fraction contained three major proteins with mol. wts. 108,000, 99,000 and 65,000.

Steroid Metabolism by the Immature Rat Testis

1974 ◽  
Vol 52 (9) ◽  
pp. 744-750 ◽  
Author(s):  
W. H. Moger ◽  
D. T. Armstrong
Keyword(s):  
Luteinizing Hormone ◽  
Male Rats ◽  
Steroid Metabolism ◽  
Immature Rat ◽  
Rat Testis ◽  
Immature Male ◽  
Immature Rats ◽  
Rapid Conversion

Treatment of hypophysectomized immature male rats with luteinizing hormone (LH) greatly increased the metabolism of both 4-[14C]progesterone and 4-[14C]testosterone by testicular homogenates. Prolactin, either alone or in combination with LH, did not influence the metabolism of either substrate. Progesterone was metabolized to 17α-hydroxyprogesterone, androstenedione, 5α-pregnan-3,20-dione, 3α-hydroxy-5α-pregnan-20-one, and 3β-hydroxy-5α-pregnan-20-one. Testosterone was metabolized to dihydrotestosterone and 5α-androstan-3α,17β-diol. On the basis of these observations it is suggested that LH stimulated the 5α-reductase(s) of the immature rat testis. Testis homogenates from immature rats with intact pituitaries were incubated with 4-[14C]3α-hydroxy-5α-pregnan-20-one. Rapid conversion to androsterone was observed, with the formation of a compound chromatographically identical with 3α, 17α-dihydroxy-5α-pregnan-20-one as an apparent intermediate. These findings demonstrate the ability of the rat testes to form androsterone from progesterone by a pathway that does not involve testosterone.

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