Bovine serum albumin , characterization

This experiment provides a nice example of the application of spectroscopy to biochemistry. After presenting the basic theory for the spectroscopic treatment of protein-ligand interactions, a procedure for characterizing the binding of methyl orange to bovine serum albumin is described. 

Specifications for chromatographic packings often describe maximum loading in terms of the maximum capacity of a unit quantity of the packing to bind some analyte, often a well characterized protein such as bovine serum albumin. The static loading capacity35 is very different from functional capacity,2 which is the maximum amount of a particular feedstock that can be loaded and still achieve acceptable purification and recovery. Functional capacity is determined empirically for each type of load and associated set... 

The imidazolate bridged Cu/Zn bimetallic complex of the cryptand (13) was structurally characterized and shown to have a Cu-Zn distance of 5.93 A (native Cu, Zn-SOD 6.2 A).146 The complex shows some activity in the dismutation of superoxide at biological pH that is retained in the presence of bovine serum albumin. 

Another derivatization approach is reduction of the hydroperoxide, followed by structural characterization of the corresponding alcohol, which is usually easier to handle. Thus, the structure of amino acid hydroperoxides can be characterized more easily if, after having ascertained the hydroperoxide nature of the compound, it is reduced to the alcohol with NaBH4. The structure of three valine hydroperoxides obtained on y-radiation of bovine serum albumin, a tripeptide (31) or valine (34) was elucidated after reduction, hydrolysis (if necessary), chromatographic separation, and application of the usual MS and NMR methods on the individual hydroxy derivatives of valine. ... 

Peters, T., Jr. and Feldhoff, R. C. 1975. Fragments of bovine serum albumin produced by limited proteolysis. Isolation and characterization of peptic fragments. Biochemistry 14, 3384-3391. 

Biophysical characterization showed that a single HRP II protein bound 17 molecules of heme [35]. In an in vitro heme polymerization assay, HRP II promoted the synthesis of hemozoin, while controls, such as the proteins bovine serum albumin and lysozyme or the homopeptides polyhistidine, polylysine, and polyasparagine, did not. FT-IR analysis of the reaction product showed the characteristic vibrations of hemozoin. The polymerization activity had a pH maximum near 4.0, which dropped off precipitously near the pKa of histidine. The heme polymerization... 

Restani, P., Ballabio, C., Cattaneo, A., Isoardi, P., Terracciano, L., and Fiocchi, A. 2004. Characterization of bovine serum albumin epitopes and their role in allergic reactions. Allergy 59, 78 21-24. 

The effects of conditioning layers of two important blood serum proteins, albumin and fibrinogen were investigated. Protein adsorption was studied using bovine serum albumin (BSA) and fibrinogen (F) from Sigma. The samples were incubated for 3 h at 37°C in solutions of albumin (1 mg/mL) and fibrinogen (0.2 mg/mL) prepared in phosphate buffered saline (PBS, 0.01 M phosphate buffer, 0.0027 M KC1, 0.137 MNaCl, pH 7.4). After the incubation period, the samples were rinsed 3 times with PBS and analyzed by the various surface characterization techniques. 

UV photolysis of CpMn(CO)3 in toluene leads to loss of CO and formation of CpMn(CO)2( ] -toluene). Kinetic studies suggest that the binding energy of the toluene is ca. 60kJmor. The binding of H2 to CpMn(CO)2 has been studied in supercritical CO2 solvent. It has been proposed that pyrylium and pyridinium salts such as (35) can be used to label proteins and thereby aid in the detection and characterization of receptor sites. Cymantrene bound to lysine residues of bovine serum albumin (BSA) has been used as a redox label. Electrochemical reduction of the label established an impressive BSA detection limit of 2 x 10 M. 

Various preparations of antisera to atropine have been reported. A racemic hemisuccinate ester was prepared and conjugated to bovine serum albumin by the carbodiimide technique. Antisera formed to the original immunogen selectively bound the R isomer (34), but a later antiserum prepared by this approach was reported to bind both R and S forms with "equal efficiency" (35). R, S-atropine was treated with diazotized p-amino-benzoic acid, and the resulting compound (which was not further characterized) was used for conjugation to bovine serum albumin by means of a carbodiimide-mediated reaction. Antisera resulting from use of this material were quite selective for the R isomer, with a cross-reaction of only about 2% for the S isomer (36). Virtanen et al. followed this procedure with S-atropine. Their antiserum bound equally to S- and R,S-atropine, as measured by displacement of tritium-labeled R,S-atropine (37). In another study (31), both racemic atropine and the S isomer were coupled to human serum albumin by the technique of Wurtzburger et al. (36), Antisera were obtained that were selective for both the R and S isomers (33). 

W.T Zhang, M.W. Capp, J.P. Bond, C.F. Anderson, M.T. Record Jr., Thermodynamic characterization of interactions of native bovine serum albumin with highly excluded (glycine betaine) and moderately... 

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