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Examples
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(C) ERAT is specific for Ser5: lack of acylation of the GAM-proghrelin mutants and preproghrelin.
PLoS ONE Alerts: New Articles Akihiko Ozawa et al. 2009
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Three mutants, namely S5A, S6A, and doubly mutated (S5A, S6A) proghrelin were generated using the Quickchange mutagenesis kit
PLoS ONE Alerts: New Articles Akihiko Ozawa et al. 2009
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(A) ERAT acylates GAM-proghrelin, but not His-tagged proghrelin, nor other precursors.
PLoS ONE Alerts: New Articles Akihiko Ozawa et al. 2009
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Treatment of His-tagged proghrelin with TEV protease to remove the His-tag has been described
PLoS ONE Alerts: New Articles Akihiko Ozawa et al. 2009
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GAM-proghrelin; lanes 5 and 6, mouse POMC; lanes 7 and 8, rat proenkephalin; lanes 9 and 10, ACTH.
PLoS ONE Alerts: New Articles Akihiko Ozawa et al. 2009
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(A) Acyltransferase activity was tested using [14C] octanoic acid transfer to either His-tagged proghrelin or GAM-proghrelin and the P2 microsomal fraction from HEL cells as an enzyme source.
PLoS ONE Alerts: New Articles Akihiko Ozawa et al. 2009
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Indeed, the specificity of ERAT was unexpected; while highly active on GAM-proghrelin, it did not acylate POMC, proenkephalin,
PLoS ONE Alerts: New Articles Akihiko Ozawa et al. 2009
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ACTH, His-tagged proghrelin, preproghrelin or a non-His-tagged proghrelin lacking the amino-terminal tripeptide GAM.
PLoS ONE Alerts: New Articles Akihiko Ozawa et al. 2009
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Aliquots of the peak fractions from Mini-S chromatography were preincubated with GAM-proghrelin and either 0.1 mM ZnCl
PLoS ONE Alerts: New Articles Akihiko Ozawa et al. 2009
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Figure 1A, left panel, shows the profile of Coomassie-stained proteins present in the reaction mixtures, while the right panel, a phosphorimage of the same gel, shows that only a modified proghrelin protein bearing the Gly-Ala-Met (GAM) amino-terminal tripeptide remaining from TEV (tobacco etch virus) protease cleavage can serve as a substrate for the reaction; amino-terminally His-tagged proghrelin was inactive as a substrate (compare lanes 2 and 4).
PLoS ONE Alerts: New Articles Akihiko Ozawa et al. 2009
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