Characterization of amyloid formation by glucagon-like peptides: role of basic residues in heparin-mediated aggregation.
Glycosaminoglycans (GAGs) have been reported to play a significant role in amyloid formation of a wide range of proteins/peptides either associated with diseases or native biological functions. The exact mechanism by which GAGs influence amyloid formation is not clearly understood. Here, we studied two closely related peptides, glucagon-like peptide 1 (GLP1) and glucagon-like peptide 2 (GLP2), for their amyloid formation in the presence and absence of the representative GAG heparin using various biophysical and computational approaches. We show that the aggregation and amyloid formation by these peptides follow distinct mechanisms: GLP1 follows nucleation-dependent aggregation, whereas GLP2 forms amyloids without any significant lag time. Investigating the role of heparin, we also found that heparin interacts with GLP1, accelerates its aggregation, and gets incorporated within its amyloid fibrils. In contrast, heparin neither affects the aggregation kinetics of GLP2 nor gets embedded within its fibrils. Furthermore, we found that heparin preferentially influences the stability of the GLP1 fibrils over GLP2 fibrils. To understand the specific nature of the interaction of heparin with GLP1 and GLP2, we performed all-atom MD simulations. Our in silico results show that the basic-nonbasic-basic (B-X-B) motif of GLP1 (K28-G29-R30) facilitates the interaction between heparin and peptide monomers. However, the absence of such a motif in GLP2 could be the reason for a significantly lower strength of interaction between GLP2 and heparin. Our study not only helps to understand the role of heparin in inducing protein aggregation but also provides insight into the nature of heparin-protein interaction.
Department of Biosciences and Bioengineering, IIT Bombay , Mumbai 400 076, India.
Narendra Nath
NN
Jha
A
A
Anoop
Srivastav
S
Ranganathan
Ganesh M
GM
Mohite
Ranjith
R
Padinhateeri
Samir K
SK
Maji
D000595:Amino Acid Sequence; D000682:Amyloid / Q000737:chemistry* / Q000648:ultrastructure; D000817:Anilino Naphthalenesulfonates / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D052216:Glucagon-Like Peptide 1 / Q000737:chemistry*; D053767:Glucagon-Like Peptide 2 / Q000737:chemistry*; D006493:Heparin / Q000737:chemistry*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007700:Kinetics; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D011485:Protein Binding; D055503:Protein Multimerization; D017433:Protein Structure, Secondary; D014364:Tryptophan / Q000737:chemistry
C515594:8-anilino-1-naphthalenesulfonic acid; D000682:Amyloid; D000817:Anilino Naphthalenesulfonates; D005456:Fluorescent Dyes; D053767:Glucagon-Like Peptide 2; D010446:Peptide Fragments; D052216:Glucagon-Like Peptide 1; D014364:Tryptophan; D006493:Heparin