Determination of 3′-phosphoadenosine-5′-phosphosulfate in cells and Golgi fractions using hydrophilic interaction liquid chromatography–mass spectrometry

Forskningsoutput: TidskriftsbidragArtikel i vetenskaplig tidskrift

Abstract

3′-Phosphoadenosine-5′-phosphosulfate (PAPS) is a key player in the sulfation of biomolecules, but methods for selective measurements are lacking. A liquid chromatography–mass spectrometry (LC–MS) approach for measuring PAPS was developed. A central feature of the method was employing hydrophilic interaction liquid chromatography (HILIC), which is highly suited for separating very polar/charged compounds, and is compatible with electrospray MS. Using simple instrumentation, the analysis time per sample was below 10 min and the method was characterized by easy sample preparation. The method was used to monitor decreasing levels of PAPS as function of sodium chlorate treatment (an inhibitor of PAPS synthesis) in whole-cell lysates as well as Golgi-fractions. The method allowed PAPS to be chromatographically separated from ADP and ATP, which can interfere with measurements if a less resolving LC–MS method is used.

Detaljer

Författare
  • Rua Kareem Dowood
  • Ravi Adusumalli
  • Emil Tykesson
  • Elin Johnsen
  • Elsa Lundanes
  • Kristian Prydz
  • Steven Ray Wilson
Enheter & grupper
Externa organisationer
  • University of Oslo
Forskningsområden

Ämnesklassifikation (UKÄ) – OBLIGATORISK

  • Cell- och molekylärbiologi

Nyckelord

Originalspråkengelska
Sidor (från-till)70-75
Antal sidor6
TidskriftJournal of Chromatography A
Volym1470
StatusPublished - 2016 okt 28
PublikationskategoriForskning
Peer review utfördJa