Add time:08/27/2019         Source:sciencedirect.com

Publisher SummaryThis chapter describes the alkylation of cysteine residues with 3-bromopropylamine, and identification by protein sequencing and quantitation by amino acid analysis. It discusses the alkylation conditions for proteins in situ on sequencer membranes that are convenient, efficient, do not require repurification of proteins before sequencing, and completely modify cysteine residues. It has been found that 3-bromopropylamine is superior to 4-VP as an on-line alkylating reagent because 4-VP has high background, and is known to cause preview. This is probably because 4-VP is more reactive than 3-bromopropylamine. Identification of cysteine is unambiguous because the PTH-APCys elutes uniquely as the last amino acid and is not affected by the ionic strength of the HPLC solvent. The chapter highlights the usefulness of 3-bromopropylamine as a reagent for quantitation of cysteine by amino acid analysis.

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