scholarly journals The N-acetylhexosaminidase components of the ram testis and epididymis

1973 ◽  
Vol 133 (3) ◽  
pp. 593-599 ◽  
Author(s):  
Sarah Bullock ◽  
Bryan Winchester
Keyword(s):  
Molecular Weight ◽  
Ion Exchange ◽  
Isoelectric Focusing ◽  
Catalytic Properties ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Multiple Forms ◽  
The Individual

Three and four N-acetylhexosaminidase components, from ram testis and epididymis respectively, have been separated by ion-exchange chromatography on DEAE-cellulose. Although they all have the same molecular weight (approx. 140000) and very similar catalytic properties towards the synthetic substrates, 4-methylumbelliferyl N-acetyl-β-glucosaminide and N-acetyl-β-galactosaminide, isoelectric focusing of the individual components showed that each had a distinct pI value. Isoelectric focusing has also been used to demonstrate the occurrence of multiple forms in ejaculated ram semen.

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.

Separation and Characterization of Two Fibrinolytic Inhibitors from Human Placenta

1971 ◽  
Vol 25 (03) ◽  
pp. 580-589 ◽  
Author(s):  
M Uszynski ◽  
U Abildgaard
Keyword(s):  
Molecular Weight ◽  
Ion Exchange ◽  
Isoelectric Focusing ◽  
Basic Protein ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Coagulation Inhibitor ◽  
Tissue Thromboplastin

SummaryProcedures for the separation of two inhibitors of the activation of plasminogen to plasmin by urokinase are described. Tissue thromboplastin was removed by adsorption to Al(0H)3 gel followed by ultracentrifugation. Plasminogen, plasminogen activator, a coagulation inhibitor and hemoglobin were removed by ion exchange chromatography (CM- or DEAE-Sephadex with NaCl gradients). The minor UK inhibitor is a relative basic protein with a pI of about 5.8. The major inhibitor was purified further by isoelectric focusing, preparative electrophoresis in polyacrylamide gel, and gel filtration. This inhibitor has α1-motility, the pI is about 5.2, and the molecular weight about 100,000. It inactivates urokinase progressively, but does not inhibit streptokinase, plasmin or thrombin.

scholarly journals Interaction between S-Type Pyocins and Microcin-II-Like Bacteriocins in Pseudomonas aeruginosa

2021 ◽  
Vol 83 (3) ◽  
pp. 72-80
Author(s):  
O.B. Balko ◽  
Keyword(s):  
Molecular Weight ◽  
Pseudomonas Aeruginosa ◽  
Ion Exchange ◽  
Protein Concentration ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Activity Increase ◽  
Activity Indices

According to our previous results, S-type bacteriocins of Pseudomonas aeruginosa are characterized by high activity against phytopathogenic Pseudomonas syringae strains. In addition to these pyocins producing strains are able to synthesize microcin-II-like bacteriocins. Presence of interaction between these two killer factors can determine methods of their use and activity increase of bacteriocins with antiphytopathogenic properties. The aim of the work was to test possibility of interaction between S-type pyocins and microcin-II-like bacteriocins of P. aeruginosa. Methods. The objects of the study were pyocins produced by 6 P. aeruginosa strains. Killer factors in composition of induced lysates were concentrated by 70% ammonium sulphate precipitation, dialyzed through dialysis membrane with molecular weight cut-off (MWCO) 3.5 kDa. Then ion-exchange chromatography with DEAE-cellulose, gel filtration with Sephadex G-75 and ultracentrifugation at 215.000 g for 1 and 4 hours were used for their separation. Protein concentration and antimicrobial activity were determined in obtained fractions. Visualization of proteins in active fraction composition was conducted by electrophoresis according to the Laemmli method. Results. Under ion-exchange chromatography with DEAE-cellulose application elution of bacteriocins available in lysate composition occurs simultaneously. The highest indices of activity and protein concentration were in the 4th fraction, containing two protein bands with molecular weight near 58 and 9 kDa, which are typical for S5 pyocin and microcin-II-like bacteriocins of P. aeruginosa. Further gel filtration of sampled fractions through Sephadex G-75 allowed to separate noted killer factors and obtaine purified fraction containing microcin-II-like pyocins only. Application of ultracentrifugation during 1 hour didn’t precipitate studied bacteriocins, whereas during 4 hours – lead to their separation. At the same time a twofold increase of activity indices for S-type pyocins in precipitates and for microcin-IIlike killer factors – in supernatants were observed. However achieved concentration was characterized by short-term effect, since in 14 days activity of supernatants decreased by 4–16 times, and for precipitates – by 80–640 times. Then revealed tendency for activity decrease continued. Conclusions. S-type pyocins and microcin-II-like bacteriocins of P. aeruginosa interact with each other, that ensures their stabilization and protects again destruction. Application of methods that cause separation of these killer factors is inexpedient, since it results into considerable decrease of bacteriocin activity indices.

scholarly journals Rat small-intestinal β-galactosidases. Studies on the fractionation of ‘acid’ β-galactosidase with isoelectric focusing, gel filtration and ion-exchange chromatography

1970 ◽  
Vol 117 (2) ◽  
pp. 369-375 ◽  
Author(s):  
Nils-Georg Asp
Keyword(s):  
Molecular Weight ◽  
Ion Exchange ◽  
High Molecular Weight ◽  
Isoelectric Focusing ◽  
Gel Filtration ◽  
Ion Exchange Resin ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Isoelectric Points ◽  
Small Intestinal

1. Different forms of the rat small-intestinal ‘acid’ β-galactosidase were separated by using the isoelectric-focusing technique. The isoelectric points of the different forms were at pH4.2, 4.6, 5.4, 6.1 and approx. 8. 2. The two forms of ‘acid’ β-galactosidase isoelectric at pH4.2 and 4.6 were completely excluded from the Sephadex G-200 gel, whereas the form isoelectric at pH8 had Kav. 0.4. The concentration and pH of the elution buffer influenced the distribution of enzyme activity between different forms. Thus, under certain conditions of ionic strength and pH, the enzyme seems to form high-molecular-weight aggregates with low isoelectric points. These may be homopolymeric aggregates or the result of binding of enzyme to, for example, membrane fragments. The forms isoelectric at pH5.4 and 6.1 are probably aggregates of intermediate size. 3. During ion-exchange chromatography at pH6.0 one fraction of ‘acid’ β-galactosidase was not retained on the column and was isoelectric at pH8 and another fraction was eluted when the buffer concentration in the eluate had increased to about 50mm. The main part of enzyme eluted in this second fraction was also isoelectric at pH8, indicating that the elution of this fraction is not a simple ion-exchange procedure but probably also involves a splitting of high-molecular-weight aggregates, originally retained because of their low isoelectric points. The enzyme subunits have a higher isoelectric point, and are therefore no longer bound to the ion-exchange resin.

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.

THE SEPARATION AND PURIFICATION OF HUMAN LUTEINIZING AND THYROTROPHIC HORMONES

1964 ◽  
Vol 29 (1) ◽  
pp. 61-69 ◽  
Author(s):  
ANNE STOCKELL HARTREE ◽  
W. R. BUTT ◽  
K. E. KIRKHAM
Keyword(s):  
Luteinizing Hormone ◽  
Ion Exchange ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Separation And Purification ◽  
Hormone Activity ◽  
Human Pituitary ◽  
Thyrotrophic Hormone

SUMMARY Ion-exchange chromatography was used to further purify a human pituitary fraction rich in thyrotrophic and luteinizing hormone activities. Approximately twofold concentration of both activities was obtained by chromatography on IRC-50 at pH 7·5, but the hormones were not separated. Subsequent chromatography on DEAE-cellulose at pH 9·5 led to a tenfold concentration of the luteinizing hormone in a fraction practically free of thyrotrophic activity and to a fourfold concentration of the thyrotrophic hormone in a fraction still exhibiting substantial luteinizing hormone activity.

scholarly journals Control of 5-aminolaevulinate synthetase activity in Rhodopseudomonas spheroides a role for trisulphides

1975 ◽  
Vol 150 (2) ◽  
pp. 245-257 ◽  
Author(s):  
J D Sandy ◽  
R C Davies ◽  
A Neuberger
Keyword(s):  
Ion Exchange ◽  
Extraction Procedure ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Synthetase Activity ◽  
Multiple Forms ◽  
Anaerobic Culture ◽  
Structural Requirement ◽  
Rhodopseudomonas Spheroides ◽  
Low Activity

1. The aminolaevulinate synthetase activator of fresh extracts of semi-anaerobically grown Rhodopseudomonas spheroids was resolved into two fractions by ion-exchange chromatography. One fraction was identified as cystine trisulphide (CySSSCy). Analysis of the other fraction indicated the presence of about equal amounts of glutathione trisulphide (GSSSG) and the mixed trisulphide of glutathione and cystine (GSSSCy). 2. Four further fractions with activator activity were observed on ion-exchange chromatography of extracts prepared by methods similar to those described earlier [Neuberger et al. (1973)Biochem. J. 136,491-499]. These activators were generated by the extraction procedure. Two of them have been identified as trisulphanedisulphonate (S5O62-) and additional cystine trisulphide. 3. For the series of polysulphanedisulphonates (-O3S-Sn-SO3-, n greater than or equal to 1), activator activity at muM concentrations was exhibited only by compounds with n greater than 3. This, together with the results described above, indicates that for a compound R-Sn-R' (where R and R' are organic or inorganic groups) the only structural requirement for activity is n greater than or equal to 3. 4. Oxygenation of a semipanaerobic culture of R. spheroids for 1.5h before harvesting the cells produced a decrease of more than 90% in the cellular content of cystine trisulphide and glutathione trisulphides. 5. Chromatography on DEAE-Sephadex confirmed the presence of multiple forms of aminolaevulinate synthetase in extracts of R. spheroides [Tuboi et al. (1970) Arch. Biochem. Biophys. 138,147-154]. Oxygenation of a semi-anaerobic culture resulted in the disappearance of high-activity enzyme (a-form) and the accumulation of low-activity enzyme (b-form) in the cell. Spontaneous activation [Marriott et al. (1969) Biochem. J. 111,385-394] And activation by cystine trisulphide both resulted in the almost complete conversion of the b-form into the a-form.

Kristallstruktur von Cs2[B6H5(SCN)]/ Crystal Structure of Cs2[B6H5(SCN)]

1998 ◽  
Vol 53 (8) ◽  
pp. 816-818 ◽  
Author(s):  
W. Preetz ◽  
S. Zander ◽  
C. Bruhn
Keyword(s):  
Crystal Structure ◽  
Ion Exchange ◽  
Structure Determination ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Orthorhombic Space Group ◽  
Space Group ◽  
X Ray

Abstract By reaction of [B6H6]2-with (SCN)2 in dichloromethane at -80 C° the thiocyanatohexaborate anion is formed and can be isolated by ion exchange chromatography on diethylaminoethyl (DEAE) cellulose. The X-ray structure determination of Cs2[B6H5(SCN)] (orthorhombic, space group Pbca with a = 9.506(5), b = 10.644(5), c = 21.857(5) Å, Z = 8) reveals that the SCN substituent is bonded via the S atom with the B-S distance of 1.885(9) Å and the B-S-C angle of 99.8(5)°. The SCN group is nearly linear (179.9(9)°).

A dipeptidylpeptidase secreted by Fasciola hepatica

Parasitology ◽  
1994 ◽  
Vol 109 (1) ◽  
pp. 113-118 ◽  
Author(s):  
C. Carmona ◽  
S. McGonigle ◽  
A. J. Dowd ◽  
A. M. Smith ◽  
S. Coughlan ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Ion Exchange ◽  
Liver Fluke ◽  
Fasciola Hepatica ◽  
Gel Filtration ◽  
Serine Proteinase ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Substrate Preference ◽  
Proteolytic Digestion

SUMMARYA dipeptidylpeptidase (DPP) was isolated from Fasciola hepatica by gel-filtration and ion-exchange chromatography. The exoproteinase is secreted by newly excysted juveniles, immature and mature flukes. The liver fluke DPP is a serine proteinase of molecular weight > 200 kDa and differs from previously characterized mammalian DPPs in its substrate preference and susceptibility to inactivation by inhibitors. The parasite DPP may function in the latter stages of the proteolytic digestion of host macromolecules. In this manner, the enzyme may be important in providing the parasite with dipeptides that could be absorbed through the intestine as nutrient.

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