Bartlet-Jones, M., Jeffery, W. A., Hansen, H. F., Pappin, D. J. C. (1995) The use of a volatile N-terminal degradation reagent for rapid, high-sensitivity sequence analysis of peptides by generation of sequence ladders. In: Techniques in Protein Chemistry. Academic Press, pp. 3-11. ISBN 1080-8914
Abstract
Publisher Summary This chapter primarily aims to explore sequencing strategies capable of rapid analysis of proteins, possibly recovered from 2D-electrophoresis gels. For this purpose, the chemistry needed to be adaptable to multiple samples and sensitive enough to work in the femtomole range. The isothiocyanate trifluoroethyl isothiocyanate (TFEITC) chemistry shows early signs of meeting these criteria. The main practical requirement is that the starting peptide is dissolved in a defined volume of coupling buffer, dependent only on the number of cycles required and the volume applied per cycle. The chapter explains, on a low-picomolar scale, that a phosphorylated tyrosine residue could be directly identified making this a potentially powerful tool for the identification of this and other sites of post-translational modification. The inherent simplicity of the process should also allow for easy automation to permit rapid processing of samples in parallel. Certain limitations are entirely instrument-related, and not relevant to the demonstration of the degradation chemistry; however, future developments in instrumentation are required to overcome these limitations.
| Item Type: | Book Section |
|---|---|
| Subjects: | bioinformatics > genomics and proteomics > design > protein network design > peptide design bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > protein tyrosine phosphatase |
| CSHL Authors: | |
| Communities: | CSHL labs > Pappin lab |
| Depositing User: | Jessica Koos |
| Date: | 1995 |
| Date Deposited: | 15 Aug 2014 18:42 |
| Last Modified: | 15 Aug 2014 18:42 |
| URI: | https://repository.cshl.edu/id/eprint/30566 |
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