B PROTEINS

1 The Colorimetric Detection and Quantitation of Total Protein B1.2 Determination of Total Nitrogen 

1 Overview of Protein Purification and Characterization B4.2 Overview of Conventional Chromatography 

1 Measurement of Functional Properties Overview of Protein Functionality Testing 

2 Measurement of Protein Hydrophobicity B5.3 Water Retention Properties of Solid Foods 

Basic Protocol 1 The Lowry Protein Assay for Determination of Total Proteins B1.1.3 Alternate Protocol 1 Modified Lowry Protein Assay for Determination 

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Fig. 3. The pH dependence, where A, B, and C represent regions corresponding to the p-K s of glutamic and aspartic acids, lysine, and argenine, respectively, of (a) protein swelling, and (b) protein acid-binding capacity. Adapted from Ref. 3.

Amino acid Three- letter code One- letter code Mass of residue in. b proteins Accessible surface area, 2 nm Hydrophobicity index ionizable side chain Occurrence in n/ proteins, /o Relative mutabihty... 

Fig. 30. Chromatographic separation of 1) acid and 2) neutral proteases (Bac. Subtilis) on Biocarb-T biosorbent. A. proteolytic activity units/ml —o— B. protein concentration (E28o) — —...

Food additives Amino acids Vitamin B Proteins (SCP)... 

Fig. 1. Spectrum of potato calluses isoperoxidases (a) non-infected plants protein extract, (b) protein extract from plants, infected by P. infestans. M - protein markers.

B) Proteins were resolved by SDS-PAGE, blotted and PG2 polypeptides detected by reaction with anti-PG2 antibodies. Lane 1, 2 pg purified PGl Lane 2, 1 pg purified PG2 Lane 3, 1 pg purified Psubunit. 

The reactions described so far do not require the involvement of the apo-B protein, neither would they necessarily result in a significant amount of protein modification. However, the peroxyl radical can attack the fatty acid to which it is attached to cause scission of the chain with the concomitant formation of aldehydes such as malondialdehyde and 4-hydroxynonenal (Esterbauer et al., 1991). Indeed, complex mixtures of aldehydes have been detected during the oxidation of LDL and it is clear that they are capable of reacting with lysine residues on the surface of the apo-B molecule to convert the molecule to a ligand for the scavenger receptor (Haberland etal., 1984 Steinbrecher et al., 1989). In addition, the lipid-derived radical may react directly with the protein to cause fragmentation and modification of amino acids. 

Wagner SL, Munoz B. Modulation of amyloid B-protein precursor processing as a means of retarding progression of Alzheimer s disease. J Clin Invest 1999 104 1329-1332. 

Winston N J 1997 Stability of cyclin B protein during meiotic maturation and the first mitotic cell division in mouse oocytes. Biol Cell 89 211-219 Winston NJ, Maro B 1995 Calmodulin-dependent protein kinase II is activated transiently in ethanol-stimulated mouse oocytes. Dev Biol 170 350-352 Winston NJ, Bourgain-Guglielmetti F, Ciemerych MA et al 2000 Early development of mouse embryos null mutant for the cyclin A2 gene occurs in the absence of maternally derived cyclin A2 gene products. Dev Biol 223 139-153... 

Gene 3 A protein Gene 8 B protein Genes 7,9 C protein Gene 6 D protein... 

Tamai et al. [30] cloned OATP-B from human brain and transporter mRNA has been detected in a number of other tissues including liver, lung, kidney, placenta, brain, heart and small intestine [26, 30]. Within the liver, OATP-B protein is local-... 

Bitan, G., Kirkitadze, M.D., Lomakin, A., Vollers, S.S., Benedek, G.B., and Teplow, D.B. (2003) Amyloid B-protein (otP) assembly ap40 and ap42 oligomerize through distinct pathways. Proc. Natl. Acad. Sci. USA 100, 330-335. 

Fig.1.5 MALDI-TOF mass spectra of purified alfalfa-derived C5-1 using (a) h uman IgG or (b) protein A. (c) Hybridoma-derived C5-1 as control. Used with permission from Ref 18.

Stable expression of the recombinant protein is another pre-requisite. For production in CHO cell culture, narrow specifications of expression level are requested by the FDA. Consequently, plant-derived proteins need to be produced according to a defined scheme and within defined specifications in order to generate material suitable for clinical trials. Therefore, the expression level must be maintained within a defined range. For example, Prodigene Inc. assessed whether the E. coli Lt-B protein was uniformly distributed throughout the defatted maize germ by analyzing samples... 

MAPK phosphatase nuclear factor-kappa B protein kinase C... 

Figure 11.11 Absorption spectrum of the protein-Coomassie brilliant blue G 250 complex A, Coomassie brilliant blue only B, protein-dye complex.

Allen, R.C. and Budowle, B., Protein Staining and Identification Techniques, Bio-Techniques Books, Natick, MA, 1999. 

Figure 1. MYST histone acetyltransferases and their associated complexes, (a) domain structure of human MYST HAT proteins, (b) Protein complexes containing MYST HAT proteins...

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