کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
7556873 | 1491298 | 2018 | 5 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
GC-MS determination of nitrous anhydrase activity of bovine and human carbonic anhydrase II and IV
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کلمات کلیدی
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: GC-MS determination of nitrous anhydrase activity of bovine and human carbonic anhydrase II and IV GC-MS determination of nitrous anhydrase activity of bovine and human carbonic anhydrase II and IV](/preview/png/7556873.png)
چکیده انگلیسی
The most widely recognized activity of the large family of the metalloenzyme carbonic anhydrases (CAs) is the diffusion-controlled hydration of CO2 to HCO3â and one proton, and the less rapid dehydration of HCO3â to CO2: CO2 + H2O â HCO3â + H+. CAs also catalyze the reaction of water with other electrophiles such as aromatic esters, sulfates and phosphates, thus contributing to lending to CAs esterase, sulfatase and phosphatase activity, respectively. Renal CAII and CAIV are involved in the reabsorption of nitrite, the autoxidation product of the signalling molecule nitric oxide (NO): 4 NO + O2 + 2 H2O â 4 ONOâ + 4 H+. Bovine and human CAII and CAIV have been reported to exert nitrite reductase and nitrous anhydride activity: 2 NO2â + 2 H+ â [2 HONO] â N2O3 + H2O. In the presence of L-cysteine, NO may be formed. In the literature, these issues are controversial, mainly due to analytical shortcomings, i.e., the inability to detect authentic HONO and N2O3. Here, we present a gas chromatography-mass spectrometry (GC-MS) assay to unambiguously detect and quantify the nitrous anhydrase activity of CAs. The assay is based on the hydrolysis of N2O3 in H218O to form ON18Oâ and 18ON18Oâ. After pentafluorobenzyl bromide derivatization and electron capture negative-ion chemical ionization of the pentafluorobenzyl nitro derivatives, quantification is performed by selected-ion monitoring of the anions with mass-to-charge (m/z) ratios of 46 (ONOâ), m/z 48 (ON18Oâ and 18ONOâ), m/z 50 (18ON18Oâ) and m/z 47 (O15NOâ, internal standard).
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Analytical Biochemistry - Volume 550, 1 June 2018, Pages 132-136
Journal: Analytical Biochemistry - Volume 550, 1 June 2018, Pages 132-136
نویسندگان
Erik Hanff, Maximilian Zinke, Anke Böhmer, Janine Niebuhr, Mirja Maassen, Volker Endeward, Norbert Maassen, Dimitrios Tsikas,