Molecular elements in FGF19 and FGF21 defining KLB/FGFR activity and specificity
Objective: To signal, FGF19 and FGF21 require co-receptor βKlotho (KLB) to act in concert with FGF receptors, and yet there is appreciable variance in the C-terminal sequences of these two novel metabolic hormones where binding is believed to be primary. We seek to determine the functional consequen...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2018-07-01
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| Series: | Molecular Metabolism |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S221287781830423X |
| _version_ | 1818231095177510912 |
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| author | Archita Agrawal Sebastian Parlee Diego Perez-Tilve Pengyun Li Jia Pan Piotr A. Mroz Ann Maria Kruse Hansen Birgitte Andersen Brian Finan Alexei Kharitonenkov Richard D. DiMarchi |
| author_facet | Archita Agrawal Sebastian Parlee Diego Perez-Tilve Pengyun Li Jia Pan Piotr A. Mroz Ann Maria Kruse Hansen Birgitte Andersen Brian Finan Alexei Kharitonenkov Richard D. DiMarchi |
| author_sort | Archita Agrawal |
| collection | DOAJ |
| description | Objective: To signal, FGF19 and FGF21 require co-receptor βKlotho (KLB) to act in concert with FGF receptors, and yet there is appreciable variance in the C-terminal sequences of these two novel metabolic hormones where binding is believed to be primary. We seek to determine the functional consequences for these amino acid differences and determine whether such information can be used to design high potency antagonists and agonists. Methods: We employed a functional in vitro assay to identify C-terminal protein fragments capable of fully blocking KLB-mediated FGF19 and 21 receptor signaling. The key residues in each hormone responsible for support full bioactivity were identified through peptide-based Ala-scanning. Chemical optimization of the peptides was employed to increase their antagonistic potency. An optimized sequence as a substituted part of a full length FGF21 was assessed for enhanced FGFR/KLB-mediated agonism using tissue culture and obese mice. Results: C-terminal FGF19 and FGF21 peptides of relatively short length were observed to potently inhibit the activity of these two hormones, in vitro and in vivo. These FGFs of different sequence also demonstrated a striking conservation of structural determinants to maintain KLB binding. A single C-terminal amino acid in FGF19 was observed to modulate relative activity through FGFR1 and FGFR4. The substitution of native FGF21 C-terminal sequence with a peptide optimized for the highest antagonistic activity resulted in significantly enhanced FGF potency, as measured by in vitro signaling and improvements in metabolic outcomes in diet-induced obese mice. Conclusions: We report here the ability of short C-terminal peptides to bind KLB and function as antagonists of FGF19 and 21 actions. These proteins maintain high conservation of sequence in those residues central to KLB binding. An FGF21 chimeric protein possessing an optimized C-terminal sequence proved to be a super-agonist in delivery of beneficial metabolic effects in obese mice. Keywords: FGF19, FGF21, KLB, FGFR isoforms, FGF antagonism, Structure-activity-relationship, Alanine-scan |
| first_indexed | 2024-12-12T10:44:56Z |
| format | Article |
| id | doaj.art-a3edd98ace354c8eb36b02867717d477 |
| institution | Directory Open Access Journal |
| issn | 2212-8778 |
| language | English |
| last_indexed | 2024-12-12T10:44:56Z |
| publishDate | 2018-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Metabolism |
| spelling | doaj.art-a3edd98ace354c8eb36b02867717d4772022-12-22T00:26:55ZengElsevierMolecular Metabolism2212-87782018-07-01134555Molecular elements in FGF19 and FGF21 defining KLB/FGFR activity and specificityArchita Agrawal0Sebastian Parlee1Diego Perez-Tilve2Pengyun Li3Jia Pan4Piotr A. Mroz5Ann Maria Kruse Hansen6Birgitte Andersen7Brian Finan8Alexei Kharitonenkov9Richard D. DiMarchi10Department of Chemistry, Indiana University, Bloomington, IN, 47405, USA; Interdisciplinary Biochemistry Graduate Program, Indiana University, Bloomington, IN, 47405, USANovo Nordisk Research Center Indianapolis, Indianapolis, IN, 46241, USAMetabolic Diseases Institute, Department of Internal Medicine, University of Cincinnati College of Medicine, Cincinnati, OH, USANovo Nordisk Research Center Indianapolis, Indianapolis, IN, 46241, USANovo Nordisk Research Center Indianapolis, Indianapolis, IN, 46241, USADepartment of Chemistry, Indiana University, Bloomington, IN, 47405, USAGlobal Research, Novo Nordisk A/S, Novo Nordisk Park, Måløv, DK-2760, DenmarkGlobal Research, Novo Nordisk A/S, Novo Nordisk Park, Måløv, DK-2760, DenmarkNovo Nordisk Research Center Indianapolis, Indianapolis, IN, 46241, USANovo Nordisk Research Center Indianapolis, Indianapolis, IN, 46241, USA; Corresponding authors.Department of Chemistry, Indiana University, Bloomington, IN, 47405, USA; Interdisciplinary Biochemistry Graduate Program, Indiana University, Bloomington, IN, 47405, USA; Novo Nordisk Research Center Indianapolis, Indianapolis, IN, 46241, USA; Corresponding authors.Objective: To signal, FGF19 and FGF21 require co-receptor βKlotho (KLB) to act in concert with FGF receptors, and yet there is appreciable variance in the C-terminal sequences of these two novel metabolic hormones where binding is believed to be primary. We seek to determine the functional consequences for these amino acid differences and determine whether such information can be used to design high potency antagonists and agonists. Methods: We employed a functional in vitro assay to identify C-terminal protein fragments capable of fully blocking KLB-mediated FGF19 and 21 receptor signaling. The key residues in each hormone responsible for support full bioactivity were identified through peptide-based Ala-scanning. Chemical optimization of the peptides was employed to increase their antagonistic potency. An optimized sequence as a substituted part of a full length FGF21 was assessed for enhanced FGFR/KLB-mediated agonism using tissue culture and obese mice. Results: C-terminal FGF19 and FGF21 peptides of relatively short length were observed to potently inhibit the activity of these two hormones, in vitro and in vivo. These FGFs of different sequence also demonstrated a striking conservation of structural determinants to maintain KLB binding. A single C-terminal amino acid in FGF19 was observed to modulate relative activity through FGFR1 and FGFR4. The substitution of native FGF21 C-terminal sequence with a peptide optimized for the highest antagonistic activity resulted in significantly enhanced FGF potency, as measured by in vitro signaling and improvements in metabolic outcomes in diet-induced obese mice. Conclusions: We report here the ability of short C-terminal peptides to bind KLB and function as antagonists of FGF19 and 21 actions. These proteins maintain high conservation of sequence in those residues central to KLB binding. An FGF21 chimeric protein possessing an optimized C-terminal sequence proved to be a super-agonist in delivery of beneficial metabolic effects in obese mice. Keywords: FGF19, FGF21, KLB, FGFR isoforms, FGF antagonism, Structure-activity-relationship, Alanine-scanhttp://www.sciencedirect.com/science/article/pii/S221287781830423X |
| spellingShingle | Archita Agrawal Sebastian Parlee Diego Perez-Tilve Pengyun Li Jia Pan Piotr A. Mroz Ann Maria Kruse Hansen Birgitte Andersen Brian Finan Alexei Kharitonenkov Richard D. DiMarchi Molecular elements in FGF19 and FGF21 defining KLB/FGFR activity and specificity Molecular Metabolism |
| title | Molecular elements in FGF19 and FGF21 defining KLB/FGFR activity and specificity |
| title_full | Molecular elements in FGF19 and FGF21 defining KLB/FGFR activity and specificity |
| title_fullStr | Molecular elements in FGF19 and FGF21 defining KLB/FGFR activity and specificity |
| title_full_unstemmed | Molecular elements in FGF19 and FGF21 defining KLB/FGFR activity and specificity |
| title_short | Molecular elements in FGF19 and FGF21 defining KLB/FGFR activity and specificity |
| title_sort | molecular elements in fgf19 and fgf21 defining klb fgfr activity and specificity |
| url | http://www.sciencedirect.com/science/article/pii/S221287781830423X |
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