Solid-phase sequencing of 32P-labeled phosphopeptides at picomole and subpicomole levels.

摘要: Publisher Summary This chapter discusses the solid-phase sequencing of 32 P-labeled phosphopeptides at picomole and subpicomole levels. Protein phosphorylation is usually restricted to hydroxyamino acids, serine, threonine, tyrosine, although histidine can occur in some systems. Specificity determinants for site-specific are provided by structural signals located within amino acid sequences adjacent site. Because sites cannot be predicted with certainty from primary structure alone, there a need methodology determine experimentally. Indirect detection methods assignment such based on novel Edman degradation products generated peptides which phosphorylserine or phosphorylthreonine groups have been specifically modified; an example this approach conversion phosphoserine residues phenylthiocarbamyl-S-ethylcysteine derivative. Such useful determination vivo sites, but require quantities excess 20 pmol.