دورية أكاديمية
Synthetic protease-activated class B GPCRs.
| العنوان: | Synthetic protease-activated class B GPCRs. |
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| المؤلفون: | Willard FS; Quantitative Biology, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN, USA. Electronic address: willardfs@lilly.com., Meredith TD; Quantitative Biology, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN, USA., Showalter AD; Diabetes and Complications, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN, USA., Ma W; Diabetes and Complications, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN, USA., Ho JD; Structural Biology, Lilly Biotechnology Center, San Diego, CA, USA., Sauder JM; Structural Biology, Lilly Biotechnology Center, San Diego, CA, USA., Sloop KW; Diabetes and Complications, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN, USA. |
| المصدر: | Biochemical and biophysical research communications [Biochem Biophys Res Commun] 2020 Sep 10; Vol. 530 (1), pp. 246-251. Date of Electronic Publication: 2020 Aug 05. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Country of Publication: United States NLM ID: 0372516 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1090-2104 (Electronic) Linking ISSN: 0006291X NLM ISO Abbreviation: Biochem Biophys Res Commun Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: <2002- >: San Diego, CA : Elsevier Original Publication: New York, Academic Press. |
| مواضيع طبية MeSH: | Exenatide/*metabolism , Glucagon-Like Peptide 1/*metabolism , Glucagon-Like Peptide-1 Receptor/*metabolism , Peptide Hydrolases/*metabolism , Protein Engineering/*methods, Animals ; Cell Line ; Exenatide/genetics ; Glucagon-Like Peptide 1/genetics ; Glucagon-Like Peptide-1 Receptor/genetics ; HEK293 Cells ; Humans ; Insulin Secretion ; Proteolysis ; Rats ; Recombinant Fusion Proteins/genetics ; Recombinant Fusion Proteins/metabolism ; Transfection |
| مستخلص: | G-protein coupled receptors (GPCRs) are the ligand detection machinery of a majority of extracellular signaling systems in metazoans. Novel chemical and biological tools to probe the structure-function relationships of GPCRs have impacted both basic and applied GPCR research. To better understand the structure-function of class B GPCRs, we generated receptor-ligand fusion chimeric proteins that can be activated by exogenous enzyme application. As a prototype, fusion proteins of the glucagon-like peptide-1 receptor (GLP-1R) with GLP-1(7-36) and exendin-4(1-39) peptides incorporating enterokinase-cleavable N-termini were generated. These receptors are predicted to generate fusion protein neo-epitopes upon proteolysis with enterokinase that are identical to the N-termini of GLP-1 agonists. This system was validated by measuring enterokinase-dependent GLP-1R mediated cAMP accumulation, and a structure-activity relationship for both linker length and peptide sequence was observed. Moreover, our results show this approach can be used in physiologically relevant cell systems, as GLP-1R-ligand chimeras were shown to induce glucose-dependent insulin secretion in insulinoma cells upon exposure to enterokinase. This approach suggests new strategies for understanding the structure-function of peptide-binding GPCRs. Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: (Copyright © 2020 Elsevier Inc. All rights reserved.) |
| فهرسة مساهمة: | Keywords: Class B; Enterokinase; G-protein coupled receptor; Glucagon-like peptide-1 receptor; Synthetic biology |
| المشرفين على المادة: | 0 (Glucagon-Like Peptide-1 Receptor) 0 (Recombinant Fusion Proteins) 89750-14-1 (Glucagon-Like Peptide 1) 9P1872D4OL (Exenatide) EC 3.4.- (Peptide Hydrolases) |
| تواريخ الأحداث: | Date Created: 20200824 Date Completed: 20210226 Latest Revision: 20210226 |
| رمز التحديث: | 20231215 |
| DOI: | 10.1016/j.bbrc.2020.07.021 |
| PMID: | 32828294 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1090-2104 |
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| DOI: | 10.1016/j.bbrc.2020.07.021 |