Abstract

The HP G1009A C-terminal sequencing system automates an efficient, reliable, and reproducible carboxy-terminal sequencing chemistry based on the introduction of diphenyl phosphoroisothiocyanatidate as the coupling reagent, pyridine as a cyclization reagent, and trimethylsilanolate as the cleavage reagent. This chapter describes the automated carboxy-terminal (C-terminal) protein sequence analysis, which enables the direct and unambiguous confirmation of the C-terminal sequence of native and expressed proteins, the detection and characterization of protein processing at the C-terminus, the identification of post-translational proteolytic cleavages, and partial sequence information on amino-terminally blocked protein samples. The sequencing methodology does not require any pre-sequencing modifications to protect side chain residues. A robust HPLC method for the identification of the thiohydantoin-amino acid derivatives, in addition to an on-line thiohydantoin-amino acid standard mixture, enables the reliable detection and identification of sequencer cycle residues. The HP G1009A sequencing system, thus, provides the flexible platform on which further developments and refinements to the chemical sequencing methodology can be accomplished.

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call

Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.