کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10532559 | 961609 | 2013 | 10 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
A sensitive fluorigenic substrate for selective in vitro and in vivo assay of leukotriene A4 hydrolase activity
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موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: A sensitive fluorigenic substrate for selective in vitro and in vivo assay of leukotriene A4 hydrolase activity A sensitive fluorigenic substrate for selective in vitro and in vivo assay of leukotriene A4 hydrolase activity](/preview/png/10532559.png)
چکیده انگلیسی
Leukotriene A4 hydrolase (LTA4H) is a bifunctional zinc-dependent metalloprotease bearing both an epoxide hydrolase, producing the pro-inflammatory LTB4 leukotriene, and an aminopeptidase activity, whose physiological relevance has long been ignored. Distinct substrates are commonly used for each activity, although none is completely satisfactory; LTA4, substrate for the hydrolase activity, is unstable and inactivates the enzyme, whereas aminoacids β-naphthylamide and para-nitroanilide, used as aminopeptidase substrates, are poor and nonselective. Based on the three-dimensional structure of LTA4H, we describe a new, specific, and high-affinity fluorigenic substrate, PL553 [l-(4-benzoyl)phenylalanyl-β-naphthylamide], with both in vitro and in vivo applications. PL553 possesses a catalytic efficiency (kcat/Km) of 3.8 ± 0.5 Ã 104 Mâ1 sâ1 using human recombinant LTA4H and a limit of detection and quantification of less than 1 to 2 ng. The PL553 assay was validated by measuring the inhibitory potency of known LTA4H inhibitors and used to characterize new specific amino-phosphinic inhibitors. The LTA4H inhibition measured with PL553 in mouse tissues, after intravenous administration of inhibitors, was also correlated with a reduction in LTB4 levels. This authenticates the assay as the first allowing the easy measurement of endogenous LTA4H activity and in vitro specific screening of new LTA4H inhibitors.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Analytical Biochemistry - Volume 441, Issue 2, 15 October 2013, Pages 152-161
Journal: Analytical Biochemistry - Volume 441, Issue 2, 15 October 2013, Pages 152-161
نویسندگان
Hervé Poras, Sophie Duquesnoy, Marie-Claude Fournié-Zaluski, Céline Ratinaud-Giraud, Bernard P. Roques, Tanja Ouimet,