کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8360257 | 1542333 | 2015 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Expression and purification of functional human glycogen synthase-1:glycogenin-1 complex in insect cells
ترجمه فارسی عنوان
بیان و تصفیه یک گلیکوژن سنتاز 1: گلیکوژنین-1 در سلول های حشرات
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کلمات کلیدی
متابولیسم انرژی، گلیکوژنز، گلوکوزیل ترانسفراز، فسفریلاسیون،
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
چکیده انگلیسی
We report the successful expression and purification of functional human muscle glycogen synthase (GYS1) in complex with human glycogenin-1 (GN1). Stoichiometric GYS1:GN1 complex was produced by co-expression of GYS1 and GN1 using a bicistronic pFastBacâ¢-Dual expression vector, followed by affinity purification and subsequent size-exclusion chromatography. Mass spectrometry analysis identified that GYS1 is phosphorylated at several well-characterised and uncharacterised Ser/Thr residues. Biochemical analysis, including activity ratio (in the absence relative to that in the presence of glucose-6-phosphate) measurement, covalently attached phosphate estimation as well as phosphatase treatment, revealed that recombinant GYS1 is substantially more heavily phosphorylated than would be observed in intact human or rodent muscle tissues. A large quantity of highly-pure stoichiometric GYS1:GN1 complex will be useful to study its structural and biochemical properties in the future, which would reveal mechanistic insights into its functional role in glycogen biosynthesis.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 108, April 2015, Pages 23-29
Journal: Protein Expression and Purification - Volume 108, April 2015, Pages 23-29
نویسندگان
Roger W. Hunter, Elton Zeqiraj, Nicholas Morrice, Frank Sicheri, Kei Sakamoto,