Bovine serum albumin, spectra

A very interesting area is the measurement of ROA in natural substances of high molecular weight. Hecht et al. (1992) showed spectra of nucleosides thymidine and 2 -desoxycytidine. Barron et al. (1992a) reported the ROA of enzymes. The same year Barron et al. (1992b) published the ROA of proteins. The L-alanine trimer, ribonuclease A, lysozyme and bovine serum albumin spectra were discussed. 

Figure 6.4. Fragmentation spectrum of a tryptic peptide obtained from bovine serum albumin. Peptide sequence LGEYGFQNALIVR, monoisotopic [M + H]+ = 1479.796, monoisotopic [M+2H]2+ =740.402. Upper panel full scan MS spectrum. Lower panel MS/MS spectrum of a doubly-charged ion at 740.7 m/z with a ladder of y ions, the distances between which correspond to amino acid residues (upper row of letters). A shorter series of b ions is also seen (lower row of letters). See Fig. 6.5 for description of nomenclature. Note the often observed phenomenon where multiply-charged ions lose the charge during fragmentation process and, therefore, have higher m/z values than the original parent ion.

Fig. 11.6. Peptide sequencing by nanoESI-CID-MS/MS from a tryptic digest of bovine serum albumin (BSA) 800 fmol of BSA were used, (a) Eull scan spectrum, (b) fragmentation of the selected doubly charged peptide ion at m/z 740.5. Adapted from Ref. [66] by permission. Nature Publishing Group, 1996.

Example The ESI mass spectrum and the charge-deconvoluted molecular weights (inset) of bovine serum albumine (BSA) as obtained from a quadrupole ion trap instrument are compared below (Fig. 11.19). Ion series A belongs to the noncovalent BSA dimer, series B results from the monomer. [24]... 

Figure 12. Effect of bovine serum albumin upon the NMR spectrum of 0.5 w/v% lyso-lecithin in 0.1M NaCl-D20 at 40°C.

We explored the detection and quantification of specific functional groups in proteins using quantitative specific chemical modifications that contain elements with high detection sensitivity. Fluorine was introduced specifically into Bovine Serum Albumin (BSA) by N-trifluoracetylation of the e-amino group of lysine using ethyl thiotrifluoracetate (47). The fluorine electron spectrum from N-trifluoracetylated BSA is shown in Figure 18. 

Figure 18. Fluorine Is spectrum from bovine serum albumin trifiuoroacetylated with ethyl thiotrifluoracetate. (Reproduced from Ref. 47. Copyright 1974, American Chemical Society.)...

Figure 8.6 clearly indicates that in the presence of free tryptophan in solution, there is no binding of TNS on the amino acid. However, in the presence of bovine serum albumin, TNS shows a fluorescence emission spectrum, indicating that TNS is bound to the protein. 

Native proteins of relatively low molecular weight show fairly well resolved spectra. Thus, Saunders et al. (7) were able to observe a spectrum of ribonuclease (molecular weight 13000) in D20 solution peaks due to at least four kinds of hydrogen could be distinguished. Other larger globular proteins, such as bovine serum albumin (molecular weight... 

Fig. 13. (A) Bovine serum albumin (1.65 mg) in 1.55 ml of 10.0 M urea solution at pH 4.15 (curve 1) after addition of 0.25 nM of NBS in 25 of water (curve 2) after addition of 50 X (0.5 /iM) of NBS (curve 3). No further decrease in ODjgo with addition of NBS was observed. (B) Human serum albumin (Hg dimer, 2.5 mg) in 3.03 ml 10 M urea solution at pH 4.15 (curve 1) after addition of 0.2 /lAf of NBS in a 1 fiM/ml solution in water (curve 2) 0.3 nM of NBS (curve 3) 0.45 nM of NBS (curve 4) 0.65 inM of NBS (curve 5) 0.75 nM of NBS (curve 6). The original spectrum was practically unchanged after the addition of only 0.1 nM of NBS. From Ramachandran and Witkop (1959).

J.E. Bruce, G.A. Anderson, J. Wen, R. Harkewicz, R.D. Smith, High-mass-measurement accuracy and 100% sequence coverage of enzymatically digested bovine serum albumin from an ESI-FTICR mass spectrum. Anal. Chem., 71 (1999) 2595. 

FIGURE 24 Positive ion ESP mass spectrum of bovine serum albumin. The average molecular mass observed here for this protein is 66,424 Da. There are two additional higher mass proteins in this sample. 

Figure 4.9 IR spectrum of deuterated Bovine Serum Albumine (BSA) at room temperature (upper spectrum). Same spectrum minus spectrum of the same sample at 116 °C (lower spectrum). Spectra are offset for clarity.

FIGURE 4.13 A MALDI-TOF mass spectrum acquired in reflectron mode showing the tryptic peptides from a digestion of bovine serum albumin. The m/z of the monoisotopic ions are assigned and can be entered into a protein database search engine to match to the theoretical trypsin digest of BSA. 

Figure 13.7 Mass spectrum of bovine serum albumin obtained on the 3-inch mass analyzer.

Proteins such as bovine serum albumin, immobilized to silica, achieve enantiomer separation primarily via hydrophobic and electrostatic interactions. Although the protein-based CSP columns have low capacity and preparative use is impossible, these phases offer the analyst the convenience of being able to resolve a broad spectrum of analytes with a single column. 

Figure 11. C Is NEXAFS spectrum of bovine serum albumin (a) and DNA (b) showing differences for the C=N bonds between 286 and 288 eV (modified from Ade at al. 1992).

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