Isolation of Fasciola hepatica haemoglobin

Parasitology ◽  
1995 ◽  
Vol 111 (2) ◽  
pp. 209-215 ◽  
Author(s):  
S. McGonigle ◽  
J. P. Dalton
Keyword(s):  
Fasciola Hepatica ◽  
Gel Filtration ◽  
Apparent Molecular Weight ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Amino Acid Sequence Analysis ◽  
Terminal Amino Acid ◽  
Terminal Amino ◽  
Haemoglobin Molecule

SUMMARYA haemoprotein released in vitro by adult Fasciola hepatica was purified by gel filtration chromatography on Sephacryl S-200 and ion-exchange chromatography on DEAE-Sepharose. The molecule, with an apparent molecular weight of > 200 kDa, contains a haem group and has absorption spectra characteristics similar to haemoglobins. N-terminal amino acid sequence analysis revealed no similarity between the F. hepatica haemoglobin and other vertebrate or invertebrate haemoglobins. Antibodies to the haemoglobin molecule can be detected in the sera of F. hepatica-infected bovines as early as 1 week after infection.

scholarly journals A cyanogen bromide fragment of β-galactosidase from Escherichia coli with α-donor activity in complementation of the enzyme from mutant M15

1976 ◽  
Vol 155 (2) ◽  
pp. 209-216 ◽  
Author(s):  
D V. Marinkovic ◽  
J N. Marinkovic
Keyword(s):  
Escherichia Coli ◽  
Gel Filtration ◽  
Exchange Chromatography ◽  
Terminal Amino Acid ◽  
E Coli ◽  
Molecular Weights ◽  
Terminal Amino ◽  
Cyanogen Bromide Fragment ◽  
Donor Activity

Aminoethylated β-galactosidase from Escherichia coli was cleaved by CNBr. The fragment C4a was purified by gel filtration and ion-exchange chromatography. The molecular weight of the fragment C4a was determined to be 9000 +/- 600. The N-terminal amino acid was found to be isoleucine. Qualitative examination of homogeneity was carried out by disc-gel electrophoresis. The fragment C4a was shown to be active as an α donor in complementation of β-galactosidase activity in vitro with E. coli mutant M15, which has a deletion in the α region of the z gene. The molecular weights of complementable fractions from mutant M15 were found to be 123 000 +/- 2500 and 507 000 +/- 11 000, and of the complemented enzyme 522 500 +/- 11 400.

scholarly journals Properties of Gluten Fractions Prepared by Ion-Exchange Chromatography on Carboxymethyl-Cellulose

1963 ◽  
Vol 16 (3) ◽  
pp. 717 ◽  
Author(s):  
JW Lee ◽  
MV Tracey ◽  
DJ Winzor ◽  
CW Wrigley ◽  
H Zentner
Keyword(s):  
Gel Electrophoresis ◽  
Wheat Gluten ◽  
Methyl Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Terminal Amino Acid ◽  
Terminal Amino ◽  
Gel Technique ◽  
Reversible Association ◽  
Carboxy Methyl

Seven fractions obtained from wheat gluten by chromatography on carboxy~ methyl-cellulose were studied by ultracentrifugation, gel electrophoresis, chemical, and N-terminal amino acid analysis. On rechromatography, five fractions eluted by sodium chloride behaved as distinct entities. illtracentrifuge experiments indicated that four of these were each undergoing rapid, reversible association. Several N-terminal amino acids were found in each of the fractions which, moreover, could be resolved by the gel technique into a number of electrophoretic bands, some bands being common to those of neighbouring fractions. Total nitrogen values showed the chromatographic samples to be essentially free from non-protein material.

THE UPTAKE OF TESTOSTERONE AND ZINC IN VITRO BY THE HUMAN BENIGN HYPERTROPHIC PROSTATE

1973 ◽  
Vol 58 (3) ◽  
pp. 405-419 ◽  
Author(s):  
M. JOAN REED ◽  
S. R. STITCH
Keyword(s):  
Ion Exchange ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Zinc Binding ◽  
Supernatant Fraction ◽  
Low Molecular Weight ◽  
Molecular Weight Peak ◽  
Steroid Binding

SUMMARY The uptake of 65Zn and [1,2-3H]testosterone by minced tissue of human benign hypertrophic prostates and the subcellular distribution of radioactivity were examined. The nature of steroid and 65Zn binding by the cytosol (105000 g supernatant) fraction was investigated by gel filtration, ion-exchange chromatography and electrophoresis. It was found that steroid binding after incubation at 4°C was specific. One or two regions of steroid binding were observed after gel filtration of the cytosol using Sephadex G-200, depending upon incubation conditions. Binding of 65Zn was found in the low molecular weight peak after G-200 gel filtration. Equimolar CdCl2 and 65ZnCl2 were incubated with [1,2-3H]testosterone and minced tissue and the cytosol was subjected to gel filtration. Compared with control values, without CdCl2, reduction of 65Zn binding by about 50% occurred, while binding of 3H-labelled steroid was unaffected. Electrophoresis and ion-exchange chromatography showed that 65Zn and 3H-labelled steroid were bound to different proteins. A sample of the zinc-binding protein was prepared by ion-exchange chromatography and the homogeneity was checked by electrophoresis.

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.

Characterisation of a Neutral Protease Produced by Micrococcus luteus

1996 ◽  
Vol 51 (5-6) ◽  
pp. 429-431 ◽  
Author(s):  
M.O. Ilori ◽  
O.O. Amund ◽  
O. Omidiji
Keyword(s):  
Molecular Weight ◽  
Citric Acid ◽  
Proteolytic Enzyme ◽  
Micrococcus Luteus ◽  
Gel Filtration ◽  
Apparent Molecular Weight ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Optimum Temperature ◽  
Optimum Ph

Abstract A proteolytic enzyme produced by a cassava-ferment­ing strain of Micrococcus luteus was extracted and puri­fied 50-fold by gel filtration and ion exchange chromatography. The optimum pH for the enzyme was 7.0, the optimum temperature 25 °C, the apparent molecular weight 42 kDa and the Km value, 0.45 mg ml-1 with casein as substrate. The enzyme was stimulated by Ca2+ and Mg2+ but inhibited by Zn2+ and Co2+ ions. Other inhibitors were EDTA, KCN, citric acid and L-cysteine indicating the enzyme to be a metalloprotease.

Purification of Clostridium perfringens type C theta toxin

1973 ◽  
Vol 19 (8) ◽  
pp. 881-885 ◽  
Author(s):  
A. H. W. Hauschild ◽  
A. Lecroisey ◽  
J. E. Alouf
Keyword(s):  
Clostridium Perfringens ◽  
Gel Filtration ◽  
Exchange Chromatography ◽  
Ammonium Sulfate Precipitation ◽  
Terminal Amino Acid ◽  
Single Protein ◽  
Dodecyl Sulfate ◽  
Type C ◽  
Terminal Amino ◽  
Anionic Polyacrylamide

Clostridium perfringens type C theta toxin was purified by ammonium sulfate precipitation, gel filtration, and ion-exchange chromatography. The purified toxin was free of alpha, beta, delta, and kappa toxins. Electrophoresis of the toxin in both the anionic polyacrylamide disc gel and in the polyacrylamide dodecyl sulfate gel yielded single protein bands. L-Lysine was determined as the sole N-terminal amino acid. The specific hemolytic activities of two purified preparations were 3.6 × 106 and 4.8 × 106 HU/mg N; the specific toxicities were 8.1 × 103 and 7.7 × 103 mouse MLD/mg N. The molecular weight determined by the polyacrylamide – dodecyl sulfate method was 74 000.

Isolation of Somatostatin (a Somatotropin Release Inhibiting Factor) of Ovine Hypothalamic Origin

1974 ◽  
Vol 52 (11) ◽  
pp. 1067-1072 ◽  
Author(s):  
P. Brazeau ◽  
W. Vale ◽  
R. Burgus ◽  
R. Guillemin
Keyword(s):  
Growth Hormone ◽  
Amino Acid ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Partition Chromatography ◽  
Inhibiting Factor ◽  
Layer Chromatography

Isolation of somatostatin, a tetradecapeptide of ovine origin inhibiting somatotropin secretion, is reported. About 490 000 hypothalamic fragments were submitted to alcohol–chloroform extraction, countercurrent distribution, ion-exchange chromatography, gel filtration, and partition chromatography. Of the 8.5 mg material thus obtained, 77% was accounted for by a peptide shown homogeneous by electrophoresis, thin-layer chromatography, and amino acid analysis. The peptide inhibits the secretion of radioimmunoassayable growth hormone at doses of ≥ 1.0 nM in vitro and 400 ng per rate in vivo.

Thermolytic digest of chicken pepsin

1981 ◽  
Vol 46 (8) ◽  
pp. 1994-2004 ◽  
Author(s):  
Miroslav Baudyš ◽  
Vladimír Kostka ◽  
Helena Keilová
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Linear Structure ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Terminal Amino Acid ◽  
Terminal Amino ◽  
Terminal Amino Acid Sequence ◽  
High Voltage Electrophoresis ◽  
Final Purification

Chicken pepsin prepared by the activation of pepsinogen was digested with thermolysin. The thermolytic digest was fractionated by chromatography on Sephadex G-25 fine. Certain fractions were subsequently subjected to ion exchange chromatography on Dowex 50-X2. The final purification was effected by paper chromatography and high voltage electrophoresis. By these procedures a series of homogeneous peptides was obtained; of the latter 54 nonoverlapping (save for a few exceptions) peptides are described in this paper. These peptides in addition to the thermolytic peptides reported before represent 80% of the linear structure of the whole molecule. The N-terminal amino acid sequence of chicken pepsin is discussed from the viewpoint of the recent data obtained by the analysis of the thermolytic digest.

Biosynthèse "In Vitro" de l'Hormone Béta-Lipotropique de Boeuf

1974 ◽  
Vol 52 (5) ◽  
pp. 349-358 ◽  
Author(s):  
X. Bertagna ◽  
M. Lis ◽  
C. Gilardeau ◽  
M. Chrétien
Keyword(s):  
Amino Acids ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Usual Method ◽  
Pituitary Hormone ◽  
Melanocyte Stimulating Hormone

Sheep beta-lipotropic hormone (β-LPH) is a pituitary hormone made of 90 amino acids and having a portion of its sequence (41–58) identical with the structure of beta-melanocyte-stimulating hormone (β-MSH). We hypothetized that β-LPH could be the biological precursor of β-MSH. We studied the biosynthesis of these two molecules by monitoring the incorporation of radioactive amino acids in beef pituitary slices. We separated β-LPH from the other radioactive proteins with the usual method of purification described previously and we characterized the proteins by ion-exchange chromatography, gel filtration, and polyacrylamide gel electrophoresis. Our results show that the pituitary slices synthesized a radioactive β-LPH which has all the characteristics of non-radioactive β-LPH. However, in the conditions used, we could not demonstrate any biosynthesis of β-MSH after 4 h incubation. These results suggest that the conversion of β-LPH into β-MSH, if it exists, is a slow process and should be studied in more prolonged incubations.

In vitro biosynthesis of γ-lipotropic hormone

1976 ◽  
Vol 54 (6) ◽  
pp. 566-570 ◽  
Author(s):  
M. Chrétien ◽  
M. Lis ◽  
C. Gilardeau ◽  
S. Benjannet
Keyword(s):  
Amino Acids ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Intermediate Compound ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Terminal Portion ◽  
In Vitro Biosynthesis ◽  
Melanophore Stimulating Hormone

Sheep γ-lipotropic hormone (γ-LPH) is a pituitary polypeptide made of 58 amino acids and is formed of the first 58 residues of β-lipotropic hormone (β-LPH). The C-terminal portion (41–58) of γ-LPH is identical with the structure of β-melanophore-stimulating hormone (β-MSH). We hypothetized in 1967 that β-LPH could be the biological precursor of β-MSH and that γ-LPH could be an intermediate compound. We demonstrated in 1974 that β-LPH is actively synthesized in the bovine pituitaries. We now studied the biosynthesis of γ-LPH by monitoring the incorporation of radioactive amino acids in beef pituitary slices. We separated γ-LPH from the other radioactive proteins with a method previously described. We characterized the radioactive proteins by ion-exchange chromatography, gel filtration and polyacrylamide gel electrophoresis. Our results show that radioactive γ-LPH was actively synthesized. This γ-LPH has all the chemical characteristics of nonradioactive γ-LPH. However, in the conditions used, we were unable to demonstrate biosynthesis of β-MSH. These results suggest that γ-LPH is biosynthesized more slowly than β-LPH and that the conversion into β-MSH, if it exists, is a slow or subactive process in the species studied.

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