A Review of: “An Introduction to Separation Science B.L. Karger,L.R. Snyder and C. Horvath; Wiley-Interscience,New York, 1973; 586 pages; hard cover; $19.50”

The preparative buffer electrofocusing of a variety of thyroid related proteins is described. The study shows that selective electrofocusing of these proteins on granulated gels can be readily achieved with buffer systems of defined chemical composition. Specific proteins were recovered in satisfactory yields (30-70%) with sample loadings in the range 25-500mg.     

A Review of: “Biological Recognition and Assembly,D. S. Eisenberg,J. A. Lake and C. F. Fox,Eds. Alan R. Liss,New York, 1980; hardbound, 355 pages, $52.00”

Two MMP-7-ase isoenzymes were purified 100-fold from rat muscle extract to apparent homogeneity, with an overall yield of 10%, using homogenization, ultracentrifugation, high-performance aqueous size-exclusion and high-performance anion exchange chromatography methods. When using a TSK G-2000SW column, the separation resulted in a 6-fold purification and 30% recovery of isoenzymes B and C. This concentrated enzyme extract was then passed through a TSK-DEAE-2SW column, using salt gradient at pH 7.5, with an additional 25-fold purification and 90% recovery of the isoenzymes. Two symmetrical enzyme peaks, representing isoenzymes B and C, were detected when performing purity tests of the active enzymes on the anion exchanger and reversed-phase HFLC columns. The procedures involved are extraction, ultracentri-fugation, chromatographies and enzyme assays and require less than five hours.     

A Review of: “RECEPTOR BIOCHEMISTRY AEiD METHODOLOGY,J.C. Venter & L.C. Harrison,Eds. Alan R. Liss,New York, 1964, hardbound; Vol.1: 238 pages, $46.00; V01.2: 306 pages, $52.00; Vo1.3: 184 pages, $34.00”

Abstract

Prothrombin complex (P.C.) preparations obtained by batch adsorption onto DEAE-Sephadex are highly enriched in C4. Based on this observation a technique has been elaborated where the P.C. analogue is further submitted to precipitation by ethanol and batch adsorption of impurities on DEAE-Cellulose. Unaltered C4 is obtained in a 2 days process with a yield corresponding to 40 % of the starting material (cryoconcentrate supernatant).     

A Review of: “Plasma Fibronectin (Structure and Function) J. McDonagh,Ed. Marcel Dekker,New York, 1985; hardbound, 269 pages, $59.75”

Abstract

Subcellular Components; Preparation and Fractionation G.D. Birnie, ed., 2nd edition, Butteworths, London and University Park Press, Baltimore, 1972; 320 pages, hardbound, $19.50

Methodological Developments in Biochemistry Volume 2 Preparative Techniques E. Reid, ed., Longman, London, 1973; 220 pages; soft cover; $9.50

Methodological Developments in Biochemistry Volume 3 Advances with Zonal Rotors E. Reid, ed., Longman, London, 1973; 273 pages; soft cover; $9.50     

A Review of: “Bioseparations, (Downstream Processing for Biotechnology) P.A. Belter,E.L. Cussler and W.S. Hu,Wiley Interscience,New York, 1988; hardbound, 368 pages, $39.95”

ABSTRACT

Covalent structural information on membrane proteins is not easily acquired since it is difficult to obtain pure membrane proteins in sufficient quantities. We have therefore examined the Bio-Rad 491 prep cell continuous elution electrophoresis apparatus as a method for providing the quantities of purified , alpha and beta subunits from (Na.K)-ATPase required for these studies. Twenty-four milligrams of crude (Na.K)-ATPase preparation was applied to the prep cell which consisted of a 7% Laemmli separating gel 4.5 cm in length. The prep cell was Y run under constant power and continuous cooling conditions. Those fractions containing the beta subunit were combined and further purified by wheat germ agglutinin affinity"' chromatography. Fractions containing the alpha subunit were combined and did not require further purification. The identity and the degree of purity of the proteins obtained using this approach was assessed utilizing SDS-PAGE, amino acid analysis 375 Copyright 1993 by Marcel Dekker, Inc. * and N-tertninal sequencing. This simple and fast method provides approximately 1.8 milligrams of each purified subunit from 24 milligrams of relatively crude microsomes. Recovery of the alpha and beta subunits from the crude (Na.K)-ATPase preparation was estimated to be 28% and 81%, respectively.     

A Review of: “Centrifugation in Biology and Medical Science,P. Sheeler,Wiley/Interscience,New York, 1981; hardbound, 269 pages, $35.00”

ABSTRACT

Peroxidases (E.C. 1.11.1.7., hydrogen donor oxidoreductase) utilize hydrogen peroxide or substituted peroxides for the oxidation of a large number of substrates. Peroxidases are widely distributed and have been isolated from many higher plants (1). The wide distribution of the enzyme suggests that it could be of great biological importance, but the physiological functions and metabolic control of these enzymes are still poorly understood. The simultaneous presence of amine oxidase and peroxidase in cell walls suggests that the peroxide generated on oxidation of the amines could be utilized by the peroxidase (2,3). Recently we have purified an amine oxidase from Hordeum vulgare (4) and we have attempted to purify the peroxidase in order to study in vitro the reconstituted coupled system. β-glucosidase (β-D-glucoside glucohydrolase E.C. 3.2.1.21.) is capable of transforming glucosides in glucose and the corresponding aglycone or disaccarides as cellobiose, sophorose, gentiobiose. This enzyme is widely distributed in plants, fungi, bacteria, yeasts and animals (5, 6). In the homogenate of Hordeum vulgare seedlings we also found β-glucosidase activity and also attempted to purify β- glucosidase. This enzyme copurified whit peroxidase up to the last step. We report here the isolation of peroxidase and β-glucosidase from Hordeum vulgare see-dlings: some molecular and kinetic properties are given.     

A Review of: “Biochemistry and Methodology of Lipids,Eds. A.R. Johnson and J.B. Davenport Wiley-Interscience,New York, 1971 x + 578 pages,ill., hard cover; $29.50”

A previous communication from this laboratory¹ as well as one from another³ described the separation of α₂-macroglobulin from swine serum. The products from both laboratories contained, in addition to α₂-macroglob-ulin, an additional macroglobulin contaminant with α₂-globulln mobility. Due to their physicochemical similarity these macroglobulins are not resolved using conventional column procedures such as ion exchange chromatography and gel filtration. Subsequent experiments have shown that immunoelectro-phoretically pure swine α₂-macroglobulln is present, in good yield (65%) in the breakthrough effluent of columns of Bio-Gel A-1.5m-Reactive Blue 2 while the contaminating macroglobulin is tightly bound. The production of highly purified swine α₂-macroglobulin utilizing this observation is the subject of the present report. The product of the separation was found to be homogeneous when subjected to Immunoelectrophoresis, at a concentration of 14–16 mg/ml, and diffused against antiswlne whole serum antibody. The production of monospecific antibody, a more stringent test for homogeneity, resulted when the purified α₂-macroglobulin was injected into rabbits. Physicochemical analyses on the purified product showed that swine and human α₂-macro-globulins are true homologs.