Quantitative Assessment of Urea In-Solution Lys-C/Trypsin Digestions Reveals Superior Performance at Room Temperature over Traditional Proteolysis at 37 °C.

Forskningsoutput: TidskriftsbidragArtikel i vetenskaplig tidskrift

Abstract

Urea-containing buffer solutions are generally used in proteomic studies to aid protein denaturation and solubilization during cell and tissue lysis. It is well-known, however, that urea can lead to carbamylation of peptides and proteins and, subsequently, incomplete digestion of proteins. By the use of cells and tissues that had been lysed with urea, different solution digestion strategies were quantitatively assessed. In comparison with traditional proteolysis at 37 °C, urea in-solution digestion performed at room temperature improved peptide and protein identification and quantitation and had a minimum impact on miscleavage rates. Furthermore, the signal intensities and the number of carbamylated and pyroglutamic acid-modified peptides decreased. Overall, this led to a reduction in the negative effects often observed for such modifications. Data are available via ProteomeXchange with identifier PXD009426.

Detaljer

Författare
Enheter & grupper
Externa organisationer
  • Tokyo Medical University
  • Skåne University Hospital
Forskningsområden

Ämnesklassifikation (UKÄ) – OBLIGATORISK

  • Klinisk medicin

Nyckelord

Originalspråkengelska
Sidor (från-till)2556−2561
Antal sidor6
TidskriftJournal of Proteome Research
Volym17
Utgivningsnummer7
StatusPublished - 2018 maj 29
PublikationskategoriForskning
Peer review utfördJa