Solid-phase sequencing of peptides and proteins

Chapter 6 Solid-Phase Sequencing of Peptides and Proteins... 

Undoubtedly, solid-phase methods have found their most important applications in the field of protein chemistry, especially for peptide synthesis. Another very important and successful application in this area is the sequencing of peptides and proteins by the solid-phase Edman degradation method. It can be said that the method is even better established than solid-phase peptide synthesis and that the uncertainties observed in solid-phase peptide synthesis do not exist in the solid-phase Edman degradation. Reviews of work in this area are available (Laursen, 1975 Laursen and Horn, 1977 Mross and Doolittle, 1977). 

The physicochemical incompatibility of the polystyrene supports with the peptides has been observed to be one of the problems associated with the original Merrifield method. This has been approached successfully by Sheppard and coworkers by the introduction of the polar polyacrylamide supports. Poly(JV-acrylylpyrrolidine) resin is another polar polymeric support developed on similar grounds. The use of these polar resin supports facilitated the solid phase synthesis of a number of peptides and protein sequences in higher yield and purity than with the polystyrene resins. However, the inadequate availability of these polar supports appears to be a limiting factor for a wider utilization at present. 

Solid-phase sequencer, an apparatus for peptide/protein sequence analysis. Automated primary structure determination of peptides and proteins using the solid-phase sequencing method is based on cova-... 

LC-MS finds wide application in the analysis of compounds that are not amenable to GC-MS, i.e. compounds that are highly polar, ionic and thermo-labile, as well as (bio)macromolecules. In environmental applications, LC-MS is applied, often in combination with off-line or on-line solid-phase extraction, to identify pesticides, herbicides, surfactants and other environmental contaminants. LC-MS plays a role in the confirmation of the presence of antibiotic residues in meat, milk and other food products. Furthermore, there is a substantial role for LC-MS in the detection and identification of new compounds in extracts from natural products and the process control of fermentation broths for industrial production of such compounds, e.g. for medicinal use. LC-MS technology is also widely applied in the characterization of peptides and proteins, e.g. rapid molecular-mass determination, peptide mapping, peptide sequencing and the study of protein conformation and noncovalent interactions of drugs, peptides and other compounds with proteins and DNA. However, the most important application area... 

Yamashiro, D., Blake, J., and Li, C. H. (1976) The use of trifluoroethanol for improved coupling in solid-phase peptide synthesis. Tetrahedron Lett. 17,1469-1472. Milton, S. C. and Milton, R. C. (1990) An improved solid-phase synthesis of a difficult-sequence peptide using hexafluoro-2-propanol. Int. J. Pept. Protein Res. 36, 193-196. 

In 2004, Kumar and Montanari introduced a fluorous tagged trivalent iodonium compound that can be used as a tag for /-Boc based solid phase peptide synthesis by tagging free amines with a perfluoroheptyl (W-C7F15) group.It is an efficient fluorous tagging reagent that has the potential to help in the synthesis of both routine and difficult peptide and protein sequences. The... 

---