Studies on γA-globulins and α2-macroglobulins. Characterization of γA- and α2-macroglobulins

1968 ◽  
Vol 46 (3) ◽  
pp. 211-216 ◽  
Author(s):  
Benjamin E. Sanders ◽  
Yang H. Oh
Keyword(s):  
Ion Exchange ◽  
Physicochemical Properties ◽  
Human Plasma ◽  
Molecular Sieve ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Normal Human Plasma ◽  
Γ Globulins ◽  
Normal Human

Fractionation into several individual components was achieved from Cohn's Fraction III of human plasma by the successive application of separation principles that depend on solubility, charge, and size (precipitation, ion-exchange chromatography, and molecular-sieve chromatography methods). Characterization was made by various electrophoretic procedures such as microzone on cellulose acetate, disc on acrylamide gel, and immunoelectrophoresis, and includes some physicochemical properties of the purified proteins. There are found to be various components of γ-globulins, α2-macroglobulins, β-glycoproteins, β-lipoproteins, and other minor proteins in Cohn's Fraction III of normal human plasma. The physicochemical properties of two γA-globulins and two α2-macroglobulins were investigated.

STUDIES ON THE NATURE OF MAMMALIAN HYPOTHALAMIC THYROTROPHIN RELEASING HORMONE USING IMMUNOCHEMICAL, CHROMATOGRAPHIC AND ENZYMIC TECHNIQUES

1975 ◽  
Vol 65 (1) ◽  
pp. 83-90 ◽  
Author(s):  
S. L. JEFFCOATE ◽  
N. WHITE
Keyword(s):  
Ion Exchange ◽  
Human Plasma ◽  
Carboxymethyl Cellulose ◽  
Mammalian Species ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Normal Human Plasma ◽  
Thyrotrophin Releasing Hormone ◽  
Releasing Hormone ◽  
Normal Human

SUMMARY Hypothalamic extracts from three mammalian species (rat, rabbit and sheep) were found to contain several ng of immunoreactive thyrotrophin releasing hormone (TRH)-like activity. This substance chromatographed on ion exchange chromatography (carboxymethyl cellulose) as a single peak that was indistinguishable from synthetic TRH. Hypothalamic TRH was also inactivated by normal human plasma at a rate (1·21–1·46%/μl plasma/h and 1·59–1·77%/50 μl plasma/min) similar to that of synthetic TRH (1·42%/μl plasma/h and 1·73%/50 μl plasma/min). This combination of chromatographic and enzymic techniques can be applied to the identification of immunoreactive TRH in body fluids.

Proteolysis in Heterocyst-Forming Cyanobacteria: Characterization of a Further Enzyme with Trypsin-Like Specificity, and of a Prolyl Endopeptidase from Anabaena variabilis

1994 ◽  
Vol 49 (1-2) ◽  
pp. 70-78 ◽  
Author(s):  
Ulrike Strohmeier ◽  
Christian Gerdes ◽  
Wolfgang Lockau
Keyword(s):  
Ion Exchange ◽  
Proteolytic Enzymes ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Anabaena Variabilis ◽  
Prolyl Endopeptidase ◽  
Cyanobacterium Anabaena

Soluble extracts of the cyanobacterium Anabaena variabilis ATCC 29413 and an engineered mutant that lacks an intracellular protease cleaving after Lys and Arg (Maldener, Lockau, Cai, and Wolk, Mol. Gen. Genet. 225, 113-120 (1991)) were separated by ion exchange chromatography, and protease profiles determined using azocasein, Nα-benzoyl-ᴅ,ʟ arginine- 4-nitroanilide and N-carbobenzoxy-glycyl-ʟ-proline-4-nitroanilide as substrates. A second enzyme cleaving at the carboxyl side of lysine and arginine, and a prolyl endopeptidase were detected, enriched and characterized. Both proteolytic enzymes appear to be located in the periplasm.

Isolation of 2-Acetamido-1-β-(L-β-Aspartamido)-1,2-Dideoxy-D-Glucose from Normal Human Urine

1972 ◽  
Vol 18 (9) ◽  
pp. 951-955 ◽  
Author(s):  
Barbara O’Neill Rowley ◽  
Paul B Hamilton
Keyword(s):  
Ion Exchange ◽  
Human Urine ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Normal Individuals ◽  
Normal Human ◽  
Elution Behavior ◽  
Solvent Systems ◽  
And Migration ◽  
Layer Chromatography

Abstract A glycopeptide was isolated from normal human urine by fractionation on a column of Sephadex G-10 and preparative ion-exchange chromatography. Elution behavior during ion-exchange chromatography in two different solvent systems, amino acids formed upon hydrolysis, and migration on high-voltage electrophoresis and thin-layer chromatography were essentially identical for this substance and for authentic 2-acetamido-l-β-(L-β-aspartamido)-1,2-dideoxy-D-glucose. A technique was developed to permit analytical-scale fractionation of individual urines followed by analysis for this glycopeptide; urine from two normal individuals contained 7 and 11 µmol of 2-acetamido-1-β-(L-β-aspartamido)-1,2-dideoxy-D-glucose per liter.

scholarly journals Isolation of albumin from whole human plasma and fractionation of albumin-depleted plasma

1976 ◽  
Vol 157 (2) ◽  
pp. 301-306 ◽  
Author(s):  
J Travis ◽  
J Bowen ◽  
D Tewksbury ◽  
D Johnson ◽  
R Pannell
Keyword(s):  
Ion Exchange ◽  
Human Plasma ◽  
Plasma Proteins ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Affinity Column ◽  
Plasma Albumin ◽  
Cibacron Blue ◽  
Blue Sepharose

The dye Cibacron Blue F-3-GA was conjugated to Sepharose to provide an affinity column for serum albumin. Passage of whole human plasma through a column of Cibacron Blue-Sepharose results in the removal of approx. 98% of the albumin. The latter can be quantitatively recovered by desorption with NaSCN. Albumin-depleted plasma can be readily resolved into discrete fractions by a combination of conventional biochemical techniques. In particular, the resolution of plasma proteins with properties similar to those of native human plasma albumin can readily be accomplished by ion-exchange chromatography of the Sepharose-dye-treated plasma on DEAE-cellulose.

scholarly journals Biochemical Characterization of a Thiol-Activated, Oxidation Stable Keratinase from Bacillus pumilus KS12

2010 ◽  
Vol 2010 ◽  
pp. 1-7 ◽  
Author(s):  
Rinky Rajput ◽  
Richa Sharma ◽  
Rani Gupta
Keyword(s):  
Ion Exchange ◽  
Bacillus Pumilus ◽  
Biochemical Characterization ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Amidolytic Activity ◽  
Alkaline Serine Protease ◽  
Sds Page ◽  
Temperature Optima

An extracellular keratinase from Bacillus pumilus KS12 was purified by DEAE ion exchange chromatography. It was a 45 kDa monomer as determined by SDS PAGE analysis. It was found to be an alkaline, serine protease with pH and temperature optima of 10 and 60C, respectively. It was thiol activated with two- and eight-fold enhancement in presence of 10 mM DTT and β-mercaptoethanol, respectively. In addition, its activity was stimulated in the presence of various surfactants, detergents, and oxidizing agents where a nearly 2- to 3-fold enhancement was observed in presence of H2O2 and NaHClO3. It hydrolyzed broad range of complex substrates including feather keratin, haemoglobin, fibrin, casein,and α-keratin. Analysis of amidolytic activity revealed that it efficiently cleaved phenylalanine → leucine → alanine- p-nitroanilides. It also cleaved insulin B chain between Val2- Asn3, Leu6-Cys7 and His10-Leu11 residues.

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