Michael B. Gosche

author keywords: caspase-3; fusion protein; protein expression; proteolysis

MeSH headings : Amino Acid Sequence; Caspase 3 / metabolism; Caspase 3 / physiology; Glutathione Transferase / genetics; Histidine / genetics; Hydrolysis; Molecular Sequence Data; Peptide Fragments / chemistry; Peptide Fragments / genetics; Peptide Fragments / metabolism; Protein Structure, Tertiary / genetics; Proteins / genetics; Proteins / isolation & purification; Proteins / metabolism; Recombinant Fusion Proteins / genetics; Recombinant Fusion Proteins / isolation & purification; Recombinant Fusion Proteins / metabolism

TL;DR: A glutathione S-transferase-fusion protein vector with a caspase-3 consensus cleavage sequence located between the N-terminal GST tag and a target protein is designed and it is shown that the enzyme efficiently cleaves the fusion protein without leaving excess amino acids on the target protein. (via Semantic Scholar)

MeSH headings : Amino Acid Sequence; Carbon Isotopes / chemistry; Caseins / analysis; Caseins / metabolism; Cell Extracts / chemistry; Cell Line, Tumor; Complex Mixtures / chemistry; Databases, Protein; Gas Chromatography-Mass Spectrometry / methods; Glass / chemistry; Humans; Isotope Labeling / methods; Molecular Sequence Data; Molecular Weight; Nitrogen Isotopes / chemistry; Peptide Fragments / analysis; Phosphopeptides / analysis; Phosphoproteins / analysis; Phosphoproteins / metabolism; Phosphoserine / chemistry; Phosphothreonine / chemistry; Photochemistry; Trypsin / chemistry; Trypsin / metabolism

TL;DR: An improved stable-isotope labeling method using a phosphoprotein isotope-coded solid-phase tag (PhIST) for isolating and measuring the relative abundances of phosphorylated peptides from complex peptide mixtures resulting from the enzymatic digestion of extracted proteins. (via Semantic Scholar)