Characterization of Proteins

Both the ease of use of this method for characterization of proteins and nucleic acids, and the abiHty to analyze many samples simultaneously for comparative purposes, have led to the prevalence of this technique. The drawbacks of a polyacrylamide matrix is that acrylamide is a neurotoxin, the reagents must be combined extremely carefiiUy, and the gels are not as pHable as most agarose gels. 

A more complete characterization of proteins was proposed in 1899 by Hausmann (130), who determined the distribution of nitrogen among amides, the basic amino acids,... 

The importance of Heinrich Ritthausen s fundamental studies, 1862 to 1899, on analytical procedures for the determination of amino acids in proteins has been emphasized in the biographical sketches which have been presented by Osborne (210), Vickery (289), and Chibnall (47). It is of particular interest to note here the prediction made by Ritt-hausen about 1870 that the amino acid composition would prove to be the most adequate basis for the characterization of proteins. Ritthausen and Kreusler (230) were the first, in 1871, to determine amino acids derived from proteins, and some of the values which they found for aspartic and glutamic acids are given in Table III (cited by Chibnall, 47, and Vickery, 286). 

Riickert, M., Wohlfarth, M., and Bringmann, G., Characterization of protein mixtures by ion-exchange chromatography coupled on-line to nuclear magnetic resonance spectroscopy, ]. Chromatogr. A, 840, 131, 1999. 

Swedberg, S. A., Characterization of protein behavior in high-performance capillary electrophoresis using a novel capillary system, Anal. Biochem., 185, 51, 1990. 

Bateman, K.R, White, R.L., Yaguchi, M., and Thibault, R, Characterization of protein glycoforms by capillary-zone electrophoresis-nanoelectrospray mass spectrometry, /. Chromatogr. A, 794, 327, 1998. 

F. Franks, Characterization of Proteins, Humana Press, Clifton, NJ, 1988. 

Evans, S. V., and MacKenzie, C. R. (1999). Characterization of protein-gly colip id recognition at the membrane biolayer. J. Molec. Recognit. 12. 

Amiri-Eliasi, B. Fenselau, C. Characterization of protein biomarkers desorbed by MALDI from whole fungal cells. Anal. Chem. 2001, 73, 5228-5231. 

B. Characterization of Proteins That Are Unstructured under Nondenaturing Conditions... 

Opiteck, G.J., Jorgenson, J.W., MacNair, J.E., Moseley, M.A., 3rd (1997). Two-dimensional SEC/RPLC coupled to mass spectrometry for the analysis of peptides. Rapid separation and characterization of protein and peptide mixtures using 1.5 microns diameter non-porous silica in packed capillary liquid chromatography/mass spectrometry. Anal. Chem. 69, 2283-2291. 

Liu B, Archer CT, Burdine L et al (2007) Label transfer chemistry for the characterization of protein—protein interactions. J Am Chem Soc 129 12348-12349... 

Usui, K., Takahashi, M., Nokihara, K. and Mihara, H. (2004b). Peptide arrays with designed alpha-helical structures for characterization of proteins from FRET fingerprint patterns. Mol. Divers. 8, 209-218. 

The significance of protein oxidation became paramount with the advent of recombinant protein biologies used as human therapeutics. Careful characterization of protein stability is essential to maintaining the efficacy of protein pharmaceuticals. If even a single side chain amino acid residue becomes oxidized, then a protein therapeutic may not have the same activity in vivo as the unmodified protein. 

About 1 in 1,000 children are born deaf, and another 1 in 1,000 develop deafness by adolescence. Although some deafness can arise during development due to external sources, much deafness in children is due to mutations in specific genes. As many forms of deafness are nonsyn-dromic — there is no other associated abnormality — identification of the responsible genes can lead to characterization of proteins that are essential for the auditory system (and often the vestibular system, as well). 

WOLF, B.P., SUMNER, L.W., SHIELDS, S.J., NIELSEN, K., GRAY, K.A., RUSSELL, D.H., Characterization of proteins utilized in the desulfurization of petroleum products by matrix-assisted laser desorption ionization time-of-flight mass spectrometry, Anal. Biochem., 1998,260,117-127. 

D. V., Characterization of protein disulphide isomerase released from activated platelets, Br. J. Haematology 90 (1995), p. 425-431... 

As one can see from Eqs. (1-5), the number of experimentally measured parameters is less than the number of unknowns independent characterization of the nine parameters in the right-hand side of Eqs. (1-5) from only five measurements is impossible. Therefore, several approaches were suggested in order to make possible characterization of protein dynamics from experimental data measured at a single resonance frequency. A low-resolution picture of protein dynamics can be obtained assuming constant values for rNH (1.02A) and 15N CSA (-160 or -170 ppm) and using various useful parametrizations for J(co) [28-31],... 

The book is separated into five major sections One short section on general aspects of spectroscopy, molecular biology and data evaluation is followed by an introduction into the NMR of commonly encountered classes of biomolecules. Thereafter, recent developments in spectroscopic techniques are highlighted. The next section describes experiments and practical aspects useful for the characterization of protein-ligand interactions. The final section presents an account on strategies for drug development using NMR written by experts from pharmaceutical industry. 

As metabolic pathways became clearer, the detailed study of the enzymes involved was facilitated by the introduction of new procedures for isolation, purification, and characterization of proteins. Developments in chromatography in the early 1940s and the introduction of gel electrophoresis allowed more efficient methods to be used to separate proteins and to analyze their primary structure, so that Sanger was able, by 1953, to report the primary structure of insulin (Chapter 10). 

Amidi M, Romeijn SG, Borchard G, Junginger HE, Hennink WE, Jiskoot W (2006) Preparation and characterization of protein-loaded N-trimethyl chi-tosan nanoparticles as nasal delivery system. J Control Release 111(1-2) 107-116. 

Refer to Section 10.7. The industry consensus is that biogenerics will be a reality in the near future, resulting from improvements in technology, which would be able to provide more precise characterization of proteins, and the continued pressure to drive down rising healthcare costs. 

Ahrer, K., and Jungbauer, A. (2006). Chromatographic and electrophoretic characterization of protein variants. /. Chromatogr. B Anal. Technol. Biomed. Life Sci. 841, 110—122. 

The CFR 21 part 11 compliant software newly released by Convergent Bioscience makes imaged cIEF a viable technique for use in QC environments for release testing of therapeutic proteins or antibody molecules. i-cIEF has also been used heavily for characterization of protein modifications such as deamidation and isomerization, as well as oligosaccharide structure analysis. ... 

Hofstadler, S. A., Wahl, J. H., Bakhtiar, R., Anderson, G. A., Bruce, J. E., and Smith, R. D. (1994). Capillary electrophoresis/Eourier-transform ion-cyclotron-resonance mass spectrometry with sustained off-resonance irradiation for the characterization of protein and peptide mixtures. ]. Am. Soc. Mass Spectrom. 5, 894 —899. 

Meyer M, Granderath K, Andreesen JR. 1995. Purification and characterization of protein PB of betaine reductase and its relationship to the corresponding proteins glycine reductase and sarcosine reductase from Eubacterium acidaminophilum. Eur J Biochem 234 184-91. 

Turner DC, Stadtman TC. 1973. Purification of protein components of the clostridial glycine reductase system and characterization of protein A as a selenoprotein. Arch Biochem Biophys 154 366-81. 

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