Using evolutionary information and ancestral sequences to understand the sequence-function relationship in GLP-1 agonists.
Glucagon-like peptide-1 (GLP-1) is an incretin hormone with therapeutic potential for type 2 diabetes. A variety of GLP-1 sequences are known from amphibian species, and some of these have been tested here and found to be able to bind and activate the human GLP-1 receptor. While little difference was observed for the in vitro potency for the human GLP-1 receptor, larger differences were found in the enzymatic stability of these peptides. Two peptides showed increased enzymatic stability, and they group together phylogenetically, though they originate from Amphibia and Reptilia. We have used ancestral sequence reconstruction to analyze the evolution of these GLP-1 molecules, including the synthesis of new peptides. We find that the increased stability could not be observed in the resurrected peptides from the common ancestor of frogs, even though they maintain the ability to activate the human GLP-1 receptor. Another method, using residue mapping on evolutionary branches yielded peptides that had maintained potency towards the receptor and also showed increased stability. This represents a new approach using evolutionary data in protein engineering.
Skovgaard
Marie
M
Novo Nordisk A/S, Novo Nordisk Park, DK-2760 Måløv, Denmark. marie.skovgaard@gmail.com
Kodra
Janos T
JT
Gram
Dorte Xenia
DX
Knudsen
Sanne Møller
SM
Madsen
Dennis
D
Liberles
David A
DA
D000595:Amino Acid Sequence; D000818:Animals; D001786:Blood Glucose / Q000378:metabolism; D002460:Cell Line; D018819:Dipeptidyl Peptidase 4 / Q000378:metabolism; D019143:Evolution, Molecular; D052216:Glucagon-Like Peptide 1 / Q000737:chemistry* / Q000235:genetics; D000067757:Glucagon-Like Peptide-1 Receptor; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D008817:Mice, Mutant Strains; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000494:pharmacology; D010802:Phylogeny; D011869:Radioligand Assay; D018027:Receptors, Glucagon / Q000819:agonists*; D013329:Structure-Activity Relationship
D001786:Blood Glucose; C000593833:GLP1R protein, human; C000593834:Glp1r protein, mouse; D000067757:Glucagon-Like Peptide-1 Receptor; D010455:Peptides; D018027:Receptors, Glucagon; D052216:Glucagon-Like Peptide 1; D018819:Dipeptidyl Peptidase 4