کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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2049973 | 1074149 | 2009 | 6 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Proof of function of a putative 3-hydroxyacyl-acyl carrier protein dehydratase from higher plants by mass spectrometry of product formation Proof of function of a putative 3-hydroxyacyl-acyl carrier protein dehydratase from higher plants by mass spectrometry of product formation](/preview/png/2049973.png)
The predicted mature portion of a putative 3-hydroxyacyl-ACP dehydratase (DH) from Arabidopsis was linked to an N-terminal poly-histidine-tag and the fusion protein expressed in Escherichia coli. Soluble dehydratase was present on induction at 25 °C and pure dehydratase eluted from a nickel-affinity column in 0.2–0.5 M imidazole. High concentrations of imidazole were necessary to retain enzyme solubility. The dehydratase reaction is reversible and 3-hydroxybutyryl- and 2-butenoyl-ACP substrates were prepared from E. coli apo-ACP. Analysis of these suggested contamination of apo-ACP with dehydratase and an additional reverse-phase chromatographic step was required during acyl carrier protein (ACP) preparation. Activity of purified dehydratase was demonstrated by mass spectrometry using 2-butenoyl-ACP, providing the first functional experimental evidence for plant DH gene sequences.
Journal: FEBS Letters - Volume 583, Issue 2, 22 January 2009, Pages 363–368