Sequence specificity of amylin-insulin interaction: a fragment-based insulin fibrillation inhibition study

Ratha, Bhisma N; Kar, Rup Kumar; Kalita, Sujan; Kalita, Sourav; Raha, Sreyan; Singha, Achintya; Garai, Kanchan; Mandal, Bhubaneswar; Bhunia, Anirban

doi:10.1016/j.bbapap.2019.01.007

Cited by 17 publications

(14 citation statements)

References 60 publications

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“…All results shown are mean ± standard deviation (SD) of three independent experiments. The observed fluorescence intensity at 482 nm for different time points was plotted against the time points, and the curve was fitted by Boltzmann equation 72,73 (eq 1)…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

“…The observed fluorescence intensity at 482 nm for different time points was plotted against the time points, and the curve was fitted by Boltzmann equation , (eq ) where A 1 is the initial fluorescence, A 2 is the maximum fluorescence, t 0 stands for the time where the fluorescence has reached to half of the maximum value, and 1/τ is the apparent rate constant of fibril growth and lag time approximated to t 0 – 2τ.…”

Section: Materials and Methodsmentioning

confidence: 99%

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Molecular Details of a Salt Bridge and Its Role in Insulin Fibrillation by NMR and Raman Spectroscopic Analysis

et al. 2020

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Insulin, a simple polypeptide hormone with huge biological importance, has long been known to self-assemble in vitro and form amyloid-like fibrillar aggregates. Utilizing highresolution NMR, Raman spectroscopy, and computational analysis, we demonstrate that the fluctuation of the carboxyl terminal (Cter) residues of the insulin B-chain plays a key role in the growth phase of insulin aggregation. By comparing the insulin sourced from bovine, human, and the modified glargine (GI), we observed reduced aggregation propensity in the GI variant, resulting from two additional Arg residues at its C-ter. NMR analysis showed atomic contacts and residue-specific interactions, particularly the salt bridge and H-bond formed among the C-ter residues Arg31 B , Lys29 B , and Glu4 A . These inter-residue interactions were reflected in strong nuclear Overhauser effects among Arg31 B δH−Glu4 A δH and Lys29 B δHs−Glu4 A δH in GI, as well as the associated downfield chemical shift of several A-chain amino terminal (N-ter) residues. The two additional Arg residues of GI, Arg31 B and Arg32 B , enhanced the stability of the GI native structure by strengthening the Arg31 B , Lys29 B , and Glu4 A salt bridge, thus reducing extensive thermal distortion and fluctuation of the terminal residues. The high stability of the salt bridge retards tertiary collapse, a crucial biochemical event for oligomerization and subsequent fibril formation. Circular dichroism and Raman spectroscopic measurement also suggest slow structural distortion in the early phase of the aggregation of GI because of the restricted mobility of the C-ter residues as explained by NMR. In addition, the structural and dynamic parameters derived from molecular dynamics simulations of insulin variants highlight the role of residue-specific contacts in aggregation and amyloid-like fibril formation.

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Section: ■ Materials and Methodsmentioning

confidence: 99%

Section: Materials and Methodsmentioning

confidence: 99%

Molecular Details of a Salt Bridge and Its Role in Insulin Fibrillation by NMR and Raman Spectroscopic Analysis

et al. 2020

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“…Insulin and amylin are both secreted by pancreatic beta cells, and there is a functional interaction between them (Ratha et al 2019). Insulin is an appetite suppressant, and its expression level was observed to be inhibited after the injection of amylin in this study, suggesting that amylin may play a regulatory role in feeding by inhibiting insulin secretion, thereby causing changes in Insulin receptor substrates, etc.…”

Section: Relationship Between Amylin and Other Appetite Factorsmentioning

confidence: 51%

Amylin Inhibits the Feeding of the Siberian Sturgeon

Wang¹,

Xu²,

Li³

et al. 2021

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Amylin is a 37-amino acid polypeptide that has been shown to be involved in feeding regulation in a few mammals, birds and goldfish. To study whether amylin regulates the feeding of Siberian sturgeon, this study cloned amylin of Siberian sturgeon and detected the distribution pattern of amylin in 15 tissues, the expression level in periprandial (pre- and post-feeding), and the changes of food intake and related appetite factors after intraperitonal injection experiment. The results showed that the expression of amylin was highest in hypothalamus, followed by duodenum, telencephalon, forebrain, midbrain, heart and liver, and low in cerebellum, pancreas, valvular intestine and other detection tissues. Compared with 1h pre-feeding, the expression level of amylin in hypothalamus was significantly increased at 1h post-feeding (P < 0.05), and the expression level of amylin in duodenum was extremely significantly increased at 1h post-feeding (P < 0.01). Compared with the control group (normal saline), the food intake at 0-1h and 1-3h after intraperitoneal injection of 100 ng/g BW and 200 ng/g BW was significantly decreased (P < 0.05), and the food intake at 3-6h in 100 ng/g BW group was extremely significantly decreased (P < 0.01), and the food intake at 3-6h in 200 ng/g BW group was significantly decreased (P < 0.05). After 1h of injection of 100 ng/g amylin, MC4R in hypothalamus was significantly increased (P < 0.05) and somatostatin was extremely significantly increased (P < 0.01), while amylin and NPY were significantly decreased (P < 0.05). CCK in valvular intestine was extremely significantly increased (P < 0.01), insulin in duodenum was significantly increased (P < 0.05), but ghrelin in duodenum had no significant change (P > 0.05). These results showed that Amylin inhibited feeding in Siberian sturgeon by down-regulating appetite stimulating factor NPY and up-regulating appetite suppressing factors somatostatin , MC4R , CCK and insulin .

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“…Bhunia and co-workers used the fibrillating core from human IAPP (N 22 FGAIL 27 , called NL6) and its derivative NFGAXL (NL6X, X = 2-aminobenzoic acid) for the inhibition of insulin amyloidogenesis. 84 The NL6 peptide was found to be more potent than NL6X. Several NOE cross peaks for E13, A14, L15, L17, E21, G23, and F25 residues of the insulin B-chain showed significant perturbations in their chemical shift upon interaction with the NL6 peptide (Figure 5C).…”

Section: Targeting the Nucleation Stepmentioning

confidence: 95%

Molecular Aspects of Insulin Aggregation and Various Therapeutic Interventions

Das

Shah

Saraogi

2022

ACS Bio Med Chem Au

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Protein aggregation leading to the formation of amyloid fibrils has various adverse effects on human health ranging from fatigue and numbness to organ failure and death in extreme cases. Insulin, a peptide hormone commonly used to treat diabetes, undergoes aggregation at the site of repeated injections in diabetic patients as well as during its industrial production and transport. The reduced bioavailability of insulin due to aggregation hinders the proper control of glucose levels in diabetic patients. Thus, it is necessary to develop rational approaches for inhibiting insulin aggregation, which in turn requires a detailed understanding of the mechanism of fibrillation. Given the relative simplicity of insulin and ease of access, insulin has also served as a model system for studying amyloids. Approaches to inhibit insulin aggregation have included the use of natural molecules, synthetic peptides or small molecules, and bacterial chaperone machinery. This review focuses on insulin aggregation with an emphasis on its mechanism, the structural features of insulin fibrils, and the reported inhibitors that act at different stages in the aggregation pathway. We discuss molecules that can serve as leads for improved inhibitors for use in commercial insulin formulations. We also discuss the aggregation propensity of fast- and slow-acting insulin biosimilars, commonly administered to diabetic patients. The development of better insulin aggregation inhibitors and insights into their mechanism of action will not only aid diabetic therapies, but also enhance our knowledge of protein amyloidosis.

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Sequence specificity of amylin-insulin interaction: a fragment-based insulin fibrillation inhibition study

Cited by 17 publications

References 60 publications

Molecular Details of a Salt Bridge and Its Role in Insulin Fibrillation by NMR and Raman Spectroscopic Analysis

Molecular Details of a Salt Bridge and Its Role in Insulin Fibrillation by NMR and Raman Spectroscopic Analysis

Amylin Inhibits the Feeding of the Siberian Sturgeon

Molecular Aspects of Insulin Aggregation and Various Therapeutic Interventions

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