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

Dowood, Rua Kareem; Adusumalli, Ravi; Tykesson, Emil LU ; Johnsen, Elin; Lundanes, Elsa; Prydz, Kristian and Wilson, Steven Ray (2016) In Journal of Chromatography A 1470. p.70-75
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... (More)

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.

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author
organization
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Contribution to journal
publication status
published
subject
keywords
3′-Phosphoadenosine-5′-phosphosulfate, Golgi, HILIC, PAPS, pHILIC, Sulfation
in
Journal of Chromatography A
volume
1470
pages
6 pages
publisher
Elsevier
external identifiers
  • scopus:84995513680
  • wos:000386983300009
ISSN
0021-9673
DOI
10.1016/j.chroma.2016.10.001
language
English
LU publication?
yes
id
f3fbba7f-a2cd-46dd-96aa-5322da3d896c
date added to LUP
2016-12-07 09:36:23
date last changed
2017-10-22 05:23:17
@article{f3fbba7f-a2cd-46dd-96aa-5322da3d896c,
  abstract     = {<p>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.</p>},
  author       = {Dowood, Rua Kareem and Adusumalli, Ravi and Tykesson, Emil and Johnsen, Elin and Lundanes, Elsa and Prydz, Kristian and Wilson, Steven Ray},
  issn         = {0021-9673},
  keyword      = {3′-Phosphoadenosine-5′-phosphosulfate,Golgi,HILIC,PAPS,pHILIC,Sulfation},
  language     = {eng},
  month        = {10},
  pages        = {70--75},
  publisher    = {Elsevier},
  series       = {Journal of Chromatography A},
  title        = {Determination of 3′-phosphoadenosine-5′-phosphosulfate in cells and Golgi fractions using hydrophilic interaction liquid chromatography–mass spectrometry},
  url          = {http://dx.doi.org/10.1016/j.chroma.2016.10.001},
  volume       = {1470},
  year         = {2016},
}