The Science of Proteins

In fact, it is a regulatory expectation that an appropriate excipient be chosen and its level (amount) in a formulation be demonstrated and justified through formulation screening and development studies (29,30). The science of protein formulation development has become increasingly sophisticated over the past 20 years and its discussion is beyond the scope of this chapter. The interested reader is referred to excellent reviews on this subject for further study (31-34). 

A crucial aspect of studying gene function is identification of the protein formed by the gene in question determination of protein function is a traditional aspect of biology. The science of protein identification and their systematic, large-scale analysis is proteomics. Protein profiles can be correlated with specific effects of disease and with alterations of function. The importance of the subject is well illustrated by the recent creation of a journal entitled Proteomics. ... 

Anfinsen C B 1973 Prinoiples that govern the folding of protein ohains Science 181 223-30... 

Pauling L, R B Corey and H R Bronson 1951. The Structure of Proteins Two Hydrogen-bonded He Configurations of the Polypeptide Chain. Proceedings of the National Academy of Sciences USA y . 211... 

CB Anfinsen. Principles that govern the folding of protein chains. Science 181 223-238, 1973. 

An aerial view of the European Synchrotron Radiation Facility at Grenoble, France, an advanced source of synchrotron x-ray radiation for use in the study of protein structure, as well as for use in the physical and material sciences. The synchrotron radiation is produced in the circular building in the lower left of the photograph. (Courtesy of ESRF.)... 

Van den Berg, G.B. Hanemajer, J.H. and Smolders, C.A., "Ultrafiltration of Protein Solutions the Role of Protein Association in Rejection and Osmotic Pressure," Journal of Membrane Science, 31 (1987) 307-320. 

Mann, M., and Wilm, M., 1995. Electro.spray ma.ss. spectrometry for protein characterization. Trends in Biochemical Sciences 20 219-224. A review of die ba.sic application of ma.ss. spectrometric methods to the analysis of protein. sequence and. structure. 

The natural world is one of eomplex mixtures petroleum may eontain 10 -10 eomponents, while it has been estimated that there are at least 150 000 different proteins in the human body. The separation methods necessary to cope with complexity of this kind are based on chromatography and electrophoresis, and it could be said that separation has been the science of the 20th century (1, 2). Indeed, separation science spans the century almost exactly. In the early 1900s, organic and natural product chemistry was dominated by synthesis and by structure determination by degradation, chemical reactions and elemental analysis distillation, liquid extraction, and especially crystallization were the separation methods available to organic chemists. 

Fig. 3-10 The 20 protein amino acids divided by R group character as (a) hydrophobic, (b) hydrophilic, and (c) mixed. (Reprinted with permission from W. K. Purves and G. H. Orians, Life The Science of Biology," pp. 63-81, Copyright 1987 by Sinauer Associates, Inc., Sunderland, MA.)...

There has been considerable and continuing investment in e-science and Grid-based computing around the world. Of particular interest for protein crystallography is the e-HTPX project funded by the UK research councils (http //www.e-htpx.ac.uk). The aim of e-HTPX is to unify the procedures of protein structure determination into a single all-encompassing interface from which users can initiate, plan, direct, and document their experiment either locally or remotely from a desktop computer. 

Methods for quantifying both the transcellular diffusion and concurrent metabolism of drugs and the unusual transcellular diffusion of membrane-interactive molecules coupled with the influence of protein binding are described in detail. To demonstrate the utility of cultured cell monolayers as a tool for basic science investigations, a subsection is devoted to the elucidation of rate-determining steps and factors in the passive diffusion of peptides across biological membranes. The chapter concludes with a discussion on the judicious use of in vitro cell monolayer results to predict in vivo results. 

Schonbranner, E.R. and Schmid, F.X. (1992) Peptidyl-prolyl cis-trans isomerase improves the efficiency of protein disulfide isomerase as a catalyst of protein folding. Proceedings of the National Academy of Sciences USA 89, 4510-4513. 

Palade, G. (1975). Intracellular aspects of the process of protein synthesis. Science 189, 347-358. 

Like FRET, today BRET is predominantly used in biological sciences, especially in the monitoring of protein-protein interactions such as hormone-receptor interaction [223, 224] and protein-DNA interaction in living systems. However, BL resonance energy transfer can also be applied in immunoassays by using for instance a peptide-tagged luciferase and a fluorescein-labeled antipeptide antibody [225]. The development of more BRET assays for small-molecule analytes is thus awaited. 

Fluorescence spectroscopy and its applications to the physical and life sciences have evolved rapidly during the past decade. The increased interest in fluorescence appears to be due to advances in time resolution, methods of data analysis and improved instrumentation. With these advances, it is now practical to perform time-resolved measurements with enough resolution to compare the results with the structural and dynamic features of macromolecules, to probe the structures of proteins, membranes, and nucleic acids, and to acquire two-dimensional microscopic images of chemical or protein distributions in cell cultures. Advances in laser and detector technology have also resulted in renewed interest in fluorescence for clinical and analytical chemistry. 

The application of the improved MS techniques presented above with highly resolving separation methods, such as 2-D electrophoresis, capillary HPLC, and CE, resulted in the creation of a new science, proteomics63 While genomics, described by DNA databases, represents the ground stage of the cell, the study of the differential status of the cell, due to various stimuli or disease states, reflects the functional expression of protein products or proteomics. Proteomics studies are aimed at identifying the proteome, the network of proteins that define the... 

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