Protein Ladder Sequencing: Towards Automation

Rong Wang, Brian T. Chait, Stephen B.H. Kent

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

6 Scopus citations

Abstract

This chapter explains protein ladder sequencing. In this method, a controlled stepwise chemical degradation is performed on one terminus of a polypeptide in the presence of a small amount of terminating reagent. The reaction yields a set of sequence-defining peptides, each differing from the next by a single amino acid residue. This protein sequencing ladder is read out rapidly in a one-step operation by matrix-assisted laser desorption mass spectrometry. Each amino acid is identified from the mass difference between successive peaks in the mass spectrum of the sequencing ladder, and the position in the data set defines the sequence of the original peptide chain. The experimental results indicate that ladder sequencing has the potential for providing extremely rapid, sensitive sequence analysis of polypeptides with high sample throughput. In addition, the method can be used to directly identify the sites and nature of post-translationally modified amino acid residues in proteins.

Original languageEnglish
Title of host publicationTechniques in Protein Chemistry
Pages19-26
Number of pages8
EditionC
DOIs
StatePublished - 1 Jan 1994
Externally publishedYes

Publication series

NameTechniques in Protein Chemistry
NumberC
Volume5
ISSN (Print)1080-8914

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