A luminescence-based assay of UDP-sugar producing pyrophosphorylases [Elektronisk resurs]
-
Decker, Daniel (författare)
-
Lindberg, Stina (författare)
-
Eriksson, Jonas (författare)
-
Kleczkowski, Leszek A. (författare)
-
Umeå universitet Teknisk-naturvetenskapliga fakulteten (utgivare)
-
Umeå universitet Teknisk-naturvetenskapliga fakulteten (utgivare)
-
Umeå universitet Teknisk-naturvetenskapliga fakulteten (utgivare)
- 2014
- Engelska.
-
Ingår i: Analytical Methods. - 1759-9660. ; 6:1, 57-61
-
Läs hela texten
-
Läs hela texten
-
Läs hela texten
-
Läs hela texten
Sammanfattning
Ämnesord
Stäng
- 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.
Ämnesord
- Natural Sciences (hsv)
- Chemical Sciences (hsv)
- Naturvetenskap (hsv)
- Kemi (hsv)
- Natural Sciences (hsv)
- Biological Sciences (hsv)
- Botany (hsv)
- Naturvetenskap (hsv)
- Biologiska vetenskaper (hsv)
- Botanik (hsv)
Inställningar
Hjälp
Beståndsinformation saknas