Split Products of Fibrinogen after Prolonged Interaction with Plasmin

1967 ◽  
Vol 18 (01/02) ◽  
pp. 089-100 ◽  
Author(s):  
J.-E Niléhn

SummaryEnd split products of fibrinogen after prolonged interaction with plasmin were separated on DEAE-cellulose. The D and E compounds measured with light ab- sorbancy constituted about 75% respectively 10% of the end split products.The D product was separated by electrophoresis into 8 components with slight difference in charge and all of them carried the same antigenic determinants. All components were found in plasmin digested fibrinogen prepared from a pool and from single donors. Four of the ∼D components were predominant. The sedimentation coefficient of the D preparation with these 4 components was 5.3 and the fraction sedimented as an homogenous peak. Slight heterogeneity of the preparation was, however, found on determination of the diffusion coefficient. The molecular weight was calculated as 80,000. On gel filtration of the D fraction on Sephadex G 200, the D products were eluted in the beginning of the albumin peak. It is suggested that the D product which is practically homogenous on ultracentrifugation dissociates on electrophoresis. The combination of the electrophoretic, immunologic and ultracentrifugal data suggests that the fibrinogen molecule consists of a series of similar repetitive units.The E product was homogenous in the ultracentrifuge and on electrophoresis. The sedimentation coefficient was 3.3 and the molecular weight was estimated to 40,800.On filtration through Sephadex G 200 the E product was hardly distinguishable from the D product and was eluted together with the ablumin.

1967 ◽  
Vol 102 (3) ◽  
pp. 763-766 ◽  
Author(s):  
J. A. W. Kirsch ◽  
R. W. Wise ◽  
I. T. Oliver

1. The foetal-specific post-albumin of rat plasma was purified by electrophoresis on a Pevicon block. 2. The sedimentation coefficient, diffusion coefficient and molecular weight were determined for this protein and found to be similar to those of adult rat plasma albumin. 3. Foetal post-albumin and adult albumin were compared immunologically and shown, with rabbit antisera, to share no antigenic determinants, suggesting different genetic control of the production of each. 4. It is suggested that the disappearance of post-albumin in the newborn animal may result from the disappearance of haemopoietic tissue from the rat liver with advancing age.


1969 ◽  
Vol 47 (8) ◽  
pp. 799-805 ◽  
Author(s):  
D. J. Ecobichon

The carboxylesterase activity of bovine liver was fractionated by DEAE-cellulose chromatography into two peaks of activity. Electrophoresis in starch gel showed that the peak eluted first was composed of a group of five electrophoretically slow bands while the second peak was a single, rapidly migrating band. Gel filtration of the crude extract on Sephadex G-100 and G-200 yielded a single peak of esterase activity containing both the electrophoretically slow and fast bands. The determination of molecular weights by gel filtration on Sephadex G-200 and G-100 yielded estimates of 52 000 and 55 000, respectively. The molecular weight estimates of the DEAE-cellulose fractionated electrophoretically slow and fast bands on Sephadex G-100 were identical, namely 55 000.


1984 ◽  
Vol 62 (5) ◽  
pp. 276-279 ◽  
Author(s):  
C. H. Lin ◽  
W. Chung ◽  
K. P. Strickland ◽  
A. J. Hudson

An isozyme of S-adenosylmethionine synthetase has been purified to homogeneity by ammonium sulfate fractionation, DEAE-cellulose column chromatography, and gel filtration on a Sephadex G-200 column. The purified enzyme is very unstable and has a molecular weight of 120 000 consisting of two identical subunits. Amino acid analysis on the purified enzyme showed glycine, glutamate, and aspartate to be the most abundant and the aromatic amino acids to be the least abundant. It possesses tripolyphosphatase activity which can be stimulated five to six times by S-adenosylmethionine (20–40 μM). The findings support the conclusion that an enzyme-bound tripolyphosphate is an obligatory intermediate in the enzymatic synthesis of S-adenosylmethionine from ATP and methionine.


1975 ◽  
Vol 39 (8) ◽  
pp. 1527-1531
Author(s):  
Zen-ichiro Hamauzu ◽  
Yukio Kamazuka ◽  
Hirokazu Kanazawa ◽  
Daizo Yonezawa

1990 ◽  
Vol 10 (2) ◽  
pp. 131-139
Author(s):  
Oyewole Adeyemo ◽  
E. O. Okegbile ◽  
O. O. Olorunsogo

For the development of immunological contraception, attention is being concentrated on the possibility of using a sperm membrane antigen. Boar sperm membrane was extracted with triton-X 100 and fractionated by Sephadex G-150 column chromatography. The glycosylated and nonglycosylated portions of protein peaks from the gel filtration were obtained by fractionating on concanavalin A-Sepharose and eluting the bound protein with 0.3 M methyl mannoside. A glycosylated fraction was found to induce sperm agglutinating antibodies in rabbit. The partially purified protein has a molecular weight of 30 kilodaltons, as determined by sodium dodecyl polyaccyrlamide gel electrophoresis. Further work is planned on the histochemical determination of the origin of this protein and species cross-activity of the antibody.


1966 ◽  
Vol 100 (3) ◽  
pp. 593-600 ◽  
Author(s):  
M Wallis ◽  
HBF Dixon

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.


1972 ◽  
Vol 130 (1) ◽  
pp. 211-219 ◽  
Author(s):  
Colin H. Self ◽  
P. David J. Weitzman

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.


1971 ◽  
Vol 124 (2) ◽  
pp. 337-343 ◽  
Author(s):  
Abraham Spector ◽  
Lu-Ku Li ◽  
Robert C. Augusteyn ◽  
Arthur Schneider ◽  
Thomas Freund

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.


1929 ◽  
Vol 12 (4) ◽  
pp. 543-554 ◽  
Author(s):  
John H. Northrop ◽  
M. L. Anson

A method is described for determining the diffusion coefficient of solutes by determining the rate of passage of the solute through a thin porous membrane between two solutions of different concentration. The method has been used to determine the diffusion coefficient of carbon monoxide hemoglobin. This was found to be 0.0420 ± 0.0005 cm.2 per day at 5°C. The molecular weight of carbon monoxide hemoglobin calculated by means of Einstein's equation from this quantity is 68,600 ± 1,000.


1970 ◽  
Vol 1 (2) ◽  
pp. 164-168
Author(s):  
Thomas M. Daniel ◽  
Lavenia E. Ferguson

Two proteins have been purified from culture filtrates of Mycobacterium tuberculosis , H 37 Ra strain by a procedure combining gel filtration, diethylaminoethyl (DEAE)-cellulose chromatography, and zone electrophoresis. The two proteins are similar in molecular weight but differ slightly in charge. The faster migrating protein, designated a 1 , is not antigenic. The slower migrating protein, designated a 2 , is antigenic both with respect to antisera and as a skin-testing antigen.


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