Peptide Maps

Protein sequencing by creation of overlap peptides (mapping),... 

M. Stromqvist, Peptide mapping using combinations of size-exclusion chromatography, reversed-phase chromatography and capillary electrophoresis , 7. Chromatogr. 667 304-310(1994). 

G. J. Opiteck, J. W. Jorgenson, M. A. Moseley III and R. J. Anderegg, Two-dimensional mia ocolumn HPLC coupled to a single-quadrupole mass spectrometer for the elucidation of sequence tags and peptide mapping , 7. Microcolumn Sep. 10 365-375 (1998). 

The use of the dynamic-FAB probe (see Section 4.4 above) has allowed the successful coupling of HPLC to this ionization technique but there is an upper limit, of around 5000 Da, to the mass of molecules which may be successfully ionized. Problem solving, therefore, often involves the use of chemical methods, such as enzymatic hydrolysis, to produce molecules of a size more appropriate for ionization, before applying techniques such as peptide mapping (see Section 5.3 below). 

Peptide mapping The process of considering the amino acid sequence information from peptides obtained by enzyme digestion in an attempt to derive the (amino acid) sequence of the parent protein. 

Reboud-Ravaux, M. Desvages, G. Chapeville, F. Irreversible inhibition and peptide mapping of urinary plasminogen activator urokinase. FEBS Lett 1982, 140, 58-62. 

Thannhauser, T. W., McWherter, C. A., and Scheraga, H. A., Peptide mapping of bovine pancreatic ribonuclease A by reverse-phase high-performance liquid chromatography. II. A two-dimensional technique for determination of disulfide pairings using a continuous-flow disulfide detection system, Anal. Biochem., 149, 322, 1985. 

Stone, K. L. and Williams, K. R., High-performance liquid chromatographic peptide mapping and amino acid analysis in the subnanomole range, /. Chromatogr., 359, 203, 1986. 

Takahashi, N., Takahashi, Y., Ishioka, N., Blumberg, B., and Putnam, F. W., Application of an automated tandem high-performance liquid chromatographic system to peptide mapping of genetic variants of human serum albumin, J. Chromatogr., 359, 181, 1986. 

Palmieri, R., Peptide mapping by capillary electrophoresis, Appl. Brief DS-774, Beckman Instruments, Fullerton, CA, 1990. 

Bonneil, E., Mercier, M. and Waldron, K.C., Reproducibility of a solid-phase trypsin microreactor for peptide mapping by capillary electrophoresis, Anal. Chim. Acta, 404, 29, 2000. 

English, R. D. Warscheid, B. Fenselau, C. Cotter, R. J. Bacillus spore identification via proteolytic peptide mapping with a miniaturized MALDITOF mass spectrometer Anal. Chem. 2003, 75, 6886-6893. 

This strategy requires a more complex system for sample processing however, it engages the advantages of improved mass accuracy and better sensitivity that derive from working with lower mass ions. Peptide maps from protein mixtures have been demonstrated to provide reliable identifications of microorganisms that are reasonably pure.79-81... 

Wagner, K., Miliotis, T., Marko-Varga, G., Bischoff, R., Unger, K.K. (2002). An automated online multidimensional HPLC system for protein and peptide mapping with integrated sample preparation. Anal. Chem. 74, 809-820. 

---