کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8302420 | 1537731 | 2014 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Recombinant PNPLA3 protein shows triglyceride hydrolase activity and its I148M mutation results in loss of function
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کلمات کلیدی
Spodoptera frugiperda 9DTTYPDSTCANickel–nitrilotriacetic acidCRBNi–NTASf9PNPLA3IFVADPNBMGYBMMYLPAATBSA - BSAbovine serum albumin - آلبومین سرم گاوAdiponutrin - آیدپوتونینtrichloroacetic acid - اسید ترشکلراکتیکbuffered methanol-complex medium - بافر متانول پیچیده محیطdithiothreitol - دیتیوتریتولbuffered glycerol-complex medium - محیط پیچیده گلیسرول بافرpatatin-like phospholipase domain containing 3 - پاتین مانند فسفولیپاز حاوی 3Pichia pastoris - پیکیا پاستوریس
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
The patatin-like phospholipase domain containing 3 (PNPLA3, also called adiponutrin, ADPN) is a membrane-bound protein highly expressed in the liver. The genetic variant I148M (rs738409) was found to be associated with progression of chronic liver disease. We aimed to establish a protein purification protocol in a yeast system (Pichia pastoris) and to examine the human PNPLA3 enzymatic activity, substrate specificity and the I148M mutation effect. hPNPLA3 148I wild type and 148M mutant cDNA were cloned into P. pastoris expression vectors. Yeast cells were grown in 3Â L fermentors. PNPLA3 protein was purified from membrane fractions by Ni-affinity chromatography. Enzymatic activity was assessed using radiolabeled substrates. Both 148I wild type and 148M mutant proteins are localized to the membrane. The wild type protein shows a predominant lipase activity with mild lysophosphatidic acid acyl transferase activity (LPAAT) and the I148M mutation results in a loss of function of both these activities. Our data show that PNPLA3 has a predominant lipase activity and I148M mutation results in a loss of function.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids - Volume 1841, Issue 4, April 2014, Pages 574-580
Journal: Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids - Volume 1841, Issue 4, April 2014, Pages 574-580
نویسندگان
Piero Pingitore, Carlo Pirazzi, Rosellina M. Mancina, Benedetta M. Motta, Cesare Indiveri, Arturo Pujia, Tiziana Montalcini, Kristina Hedfalk, Stefano Romeo,