title: Determination of 3'-phosphoadenosine-5'-phosphosulfate in cells and Golgi fractions using hydrophilic interaction liquid chromatography-mass spectrometry.
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 10min 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.
authors.affiliation: Department of Chemistry, University of Oslo, Post Box 1033, Blindern, NO-0315 Oslo, Norway.
authors.forename: Rua Kareem
authors.initials: RK
authors.lastname: Dowood
authors.affiliation: Department of Biosciences, University of Oslo, Post Box 1066, Blindern, NO-0316 Oslo, Norway.
authors.forename: Ravi
authors.initials: R
authors.lastname: Adusumalli
authors.affiliation: Department of Experimental Medical Science, Lund University, Box 117, 221 00, Lund, Sweden.
authors.forename: Emil
authors.initials: E
authors.lastname: Tykesson
authors.affiliation: Department of Chemistry, University of Oslo, Post Box 1033, Blindern, NO-0315 Oslo, Norway.
authors.forename: Elin
authors.initials: E
authors.lastname: Johnsen
authors.affiliation: Department of Chemistry, University of Oslo, Post Box 1033, Blindern, NO-0315 Oslo, Norway.
authors.forename: Elsa
authors.initials: E
authors.lastname: Lundanes
authors.affiliation: Department of Biosciences, University of Oslo, Post Box 1066, Blindern, NO-0316 Oslo, Norway.
authors.forename: Kristian
authors.initials: K
authors.lastname: Prydz
authors.affiliation: Department of Chemistry, University of Oslo, Post Box 1033, Blindern, NO-0315 Oslo, Norway. Electronic address: stevenw@kjemi.uio.no.
authors.forename: Steven Ray
authors.initials: SR
authors.lastname: Wilson
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods; D004285:Dogs; D006056:Golgi Apparatus / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D061985:Madin Darby Canine Kidney Cells; D010724:Phosphoadenosine Phosphosulfate / Q000032:analysis*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D010724:Phosphoadenosine Phosphosulfate
keywords: 3′-Phosphoadenosine-5′-phosphosulfate; Golgi; HILIC; PAPS; Sulfation; pHILIC