Reversed-phase liquid chromatography of peptides for direct micro-sequencing.
Tryptic and cyanogen bromide peptides derived from yeast aspartyl-tRNA synthetase and from Escherichia coli ribosomal proteins were separated by reversed-phase liquid chromatography, employing volatile buffers of low ionic strength. The conditions used allow the performance of micro-sequencing without desalting or extensive lyophilization, and can therefore be applied to peptide mixtures containing hydrophobic fragments which tend to precipitate. To prevent losses of peptides, direct ultra-violet detection of the peptides was preferred, to detection by post-column derivatization with an additional stream splitting device. Preparative separations were performed with 5-10 nmol of peptide mixture; analytical runs were made with 5-10 micrograms of protein hydrolysate.
D000595:Amino Acid Sequence; D001226:Aspartate-tRNA Ligase / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003488:Cyanogen Bromide; D004926:Escherichia coli / Q000032:analysis; D010446:Peptide Fragments / Q000302:isolation & purification*; D012269:Ribosomal Proteins / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D014357:Trypsin
D001426:Bacterial Proteins; D010446:Peptide Fragments; D012269:Ribosomal Proteins; D014357:Trypsin; D001226:Aspartate-tRNA Ligase; D003488:Cyanogen Bromide