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A luminescence-base...
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Decker, DanielUmeå universitet,Institutionen för fysiologisk botanik,Umeå Plant Science Centre (UPSC)
(författare)
A luminescence-based assay of UDP-sugar producing pyrophosphorylases
- Artikel/kapitelEngelska2014
Förlag, utgivningsår, omfång ...
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2014
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electronicrdacarrier
Nummerbeteckningar
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LIBRIS-ID:oai:DiVA.org:umu-85622
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https://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-85622URI
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https://doi.org/10.1039/C3AY41811aDOI
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Språk:engelska
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Sammanfattning på:engelska
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Ämneskategori:ref swepub-contenttype
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Ämneskategori:art swepub-publicationtype
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A coupled luminescence assay was applied to monitor pyrophosphate (PPi) production by either purified barley UDP-glucose pyrophosphorylase (UGPase) or purified Leishmania UDP-sugar pyrophosphorylase (USPase). In the assay, the PPi produced by the pyrophosphorylases was converted to ATP by ATP-sulfurylase, and the ATP produced was linked to luminescent light formation through the action of firefly luciferase. The assay allowed for a quantitative measurement of UGPase and USPase activities, down to a pmol per min level. The activities were linear with time and proportional to the amount of the enzyme added, and were neither affected by Pi nor by DTT. For UGPase, K-m values with UTP and Glc-1-P were 0.14 and 0.26 mM, respectively, whereas for USPase the respective K-m values with UTP, Glc-1-P and Gal-1-P were 0.4, 2.9 and 3.9 mM. Possible applications of the luminescence-based assay for not only UDP-sugar producing pyrophosphorylases, but also other types of pyrophosphorylases are discussed.
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Biuppslag (personer, institutioner, konferenser, titlar ...)
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Lindberg, StinaUmeå universitet,Kemiska institutionen(Swepub:umu)stabed97
(författare)
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Eriksson, JonasUmeå universitet,Kemiska institutionen(Swepub:umu)joer0061
(författare)
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Kleczkowski, Leszek A.Umeå universitet,Institutionen för fysiologisk botanik,Umeå Plant Science Centre (UPSC)(Swepub:umu)lekl0001
(författare)
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Umeå universitetInstitutionen för fysiologisk botanik
(creator_code:org_t)
Sammanhörande titlar
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Ingår i:Analytical Methods6:1, s. 57-611759-96601759-9679
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