Albumin crystalline

Reagents. Two synthetic peptides are required—for example, the peptides described previously (Melchior et al, 1993) work well in our hands. The NLS peptide sequence is CGGGPKKKRKVED, and the control, nonfunctional peptide is the reverse NLS sequence, CGGGDEVKRKKKP. Conjugation to the carrier protein is through the N-terminal cysteine residue and the glycine residues serve as a flexible hinge or spacer to separate the peptide from the protein itself. Bovine serum albumin (crystalline)... 

Trichloroethanoic acid, CCI3COOH. A crystalline solid which rapidly absorbs water vapour m.p. 58°C, b.p. 196-5" C. Manufactured by the action of chlorine on ethanoic acid at 160°C in the presence of red phosphorus, sulphur or iodine. It is decomposed into chloroform and carbon dioxide by boiling water. It is a much stronger acid than either the mono- or the dichloro-acids and has been used to extract alkaloids and ascorbic acid from plant and animal tissues. It is a precipitant for proteins and may be used to test for the presence of albumin in urine. The sodium salt is used as a selective weedkiller. 

Sorensen and Haugaard147 showed that by a suitable fractionation of crystalline albumin there could be obtained a series of fractions which, although crystalline, varied widely in their properties, particularly in regard to their carbohydrate contents. The most soluble fraction contained about fifty times as much carbohydrate as the least soluble. In most of these studies the carbohydrates were estimated by modification of the orcinol colorimetric method of Tillmans and Philipi181 which could... 

According to Flory (10), the concept that proteins and carbohydrates are polymeric goes back to at least the work of Hlasiwetz and Habermann (11). In 1871 they proposed that these substances were made up of a number of species differing from one another with respect to the degree of molecular condensation. Flory also noted that Hlasiwetz and Habermann differentiate "soluble and unorganized" members of these substances, for example dextrin and albumin, from "insoluble organized" members, such as cellulose or keratin. This distinction is the precursor of the present day differentiation between non-crystalline and crystalline polymers. 

Lactose.—The whey is concentrated to a small volume on a ring burner which causes almost complete precipitation of the albumin. The mixture is filtered through linen, and the filtrate is again concentrated until lactose separates. When the mixture is cold the crude crystalline sugar is collected by filtration through linen on a Buchner funnel and dried. A second portion of lactose is obtained by further concentration of the mother liquor—this time on the water bath. Yield of crude material 70-75 g. 

The protein used was crystalline bovine serum albumin (BSA) obtained from Armour Laboratories. Sodium dodecyl sulfate (SDS) was synthesized from pure dodecyl alcohol and chlorosulfonic acid, recrystallized, and washed with ethyl ether. Sulfuric acid was reagent grade purchased from the J. T. Baker Chemical Co. and used directly. Reagent grade potassium hydroxide purchased from the Mallinkrodt Co. was purified by foaming a concentrated solution, removing the foam, and using the solution directly. 

Tphe complexing of virtually all purines with aromatic molecules seems - to have far-reaching biological significance. For example, it is known that caffeine affects the rates of many enzymatic reactions (e.g., 0.01, 0.05, and 0.10M caffeine will inhibit salivary amylase 29, 54, and 72% respectively) (12), and purine can decrease the helix-coil transition temperature of the proteins bovine serum albumin and lysozyme (2). It is not unreasonable to expect the involvement of caffeine-aromatic and purine-aromatic complexes because caffeine derivatives and purine complex with the aromatic amino acids tyrosine, phenylalanine, and tryptophan (2). (In fact tryptophan forms a stable 1 to 1 crystalline complex in 0.5M theophylline glycol.)... 

Protein was measured by the method of Lowry eit al (64). Crystalline bovine serum albumin (Sigma Chemical Company) served as a standard. 

The assays for the enzyme synthesis of sulfogalactosyl-glycerolipid, sulfatides and galactocerebrosides were carried out as previously described respectively by Subba Rao, et al. (28) Sarlieve, et al. 05, 29), and Neskovic, et al. (30). The assay for 2, 3 cyclic nucleotide phosphohydrolase was performed according to the method of Prohaska, et al. (31). EL coli. alkaline phosphatase type III-S, 2, 3 -cAMP, and sodium deoxycholate were obtained from Sigma (St. Louis, Mo.). Protein was determined by the method of Lowry, et al. (32) with crystalline bovine serum albumin as the standard. 

Free fatty acids, derived primarily from adipocyte triglycerides, are transported as a physical complex with plasma albumin. Triglycerides and cholesteryl esters are transported in the core of plasma lipoproteins [134], Deliconstantinos observed the physical state of the Na+/K+-ATPase lipid microenvironment as it changed from a liquid-crystalline form to a gel phase [135], The studies concerning the albumin-cholesterol complex, its behavior, and its role in the structure of biomembranes provided important new clues as to the role of this fascinating molecule in normal and pathological states [135]. 

In our experiments crystalline bovine serum albumin from Fluka, cat.no. 05470, (FLUKA albumin) was used for calibration and traceability purposes. The Fluka product was tested for traceability against the National Institute of Standards and Technology (NIST), USA Standard Reference Material (SRM) 927c (Total Protein Standard) bovine serum albumin (NIST albumin). Two different statistical techniques were used to evaluate traceability of the FFUKA albumin to the NIST albumin. 

It has been observed that acidic drugs exhibit strong binding affinities for the serum albumins. Strong CD is induced in sulfaethidole when this compound binds to either crystalline or fraction V bovine serum albumin [68]. The data... 

Analytical methods. Chlorophyll was estimated after extraction into 80% (v/v) acetone according to Amon et al., (1974). Protein was determined according to Wang and Smith (1975) using crystalline bovine serum albumin as standard. 

Proteins. Crystalline lysozyme, bovine serum albumin, ribonucle-ase,and pepsin were obtained from the Armour Laboratories and crystalline /3-lactoglobulin was prepared from skimmed milk. a-Lactalbu-min was obtained from W. G. Gordon of this laboratory. Crystalline ovalbumin and human serum albumin were obtained from Nutritional Biochemical Corp. and purified pigskin gelatin from the Eastman Kodak Co. 

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