کد مقاله کد نشریه سال انتشار مقاله انگلیسی ترجمه فارسی نسخه تمام متن
16902 42621 2016 5 صفحه PDF ندارد دانلود کنید
عنوان انگلیسی مقاله
Linker engineering for fusion protein construction: Improvement and characterization of a GLP-1 fusion protein
ترجمه فارسی عنوان
مهندسی لینکر برای ساخت پروتئین فیوژن: بهبود و مشخص کردن یک پروتئین فیوژن GLP-1
کلمات کلیدی
GLP-1؛ آديپونکتين؛ مهندسی لینکر؛ دیابت
موضوعات مرتبط
مهندسی و علوم پایه مهندسی شیمی بیو مهندسی (مهندسی زیستی)
چکیده انگلیسی


• Different linker peptides were introduced into the fusion protein construction.
• The optimized fusion protein showed high potency in receptors activation.
• The optimized fusion protein showed high potency in glucose- and lipid-lowering.

Protein engineering has been successfully applied in protein drug discovery. Using this technology, we previously have constructed a fusion protein by linking the globular domain of adiponectin to the C-terminus of a glucagon-like peptide-1 (GLP-1) analog. Herein, to further improve its bioactivity, we reconstructed this fusion protein by introducing linker peptides of different length and flexibility. The reconstructed fusion proteins were overexpressed in Escherichia coli and purified using nickel affinity chromatography. Their agonist activity towards receptors of GLP-1 and adiponectin were assessed in vitro by using luciferase assay and AMP-activated protein kinase (AMPK) immunoblotting, respectively. The effects of the selected fusion protein on glucose and lipid metabolism were evaluated in mice. The fusion protein reconstructed using a linker peptide of AMGPSSGAPGGGGS showed high potency in activating GLP-1 receptor and triggering AMPK phosphorylation via activating the adiponectin receptor. Remarkably, the optimized fusion protein was highly effective in lowering blood glucose and lipids in mice. Collectively, these findings demonstrate that the bioactivity of this GLP-1 fusion protein can be significantly promoted by linker engineering, and indicate that the optimized GLP-1 fusion protein is a promising lead structure for anti-diabetic drug discovery.

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ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Enzyme and Microbial Technology - Volume 82, January 2016, Pages 105–109
نویسندگان
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