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Butyrylcholinesterase interactions with amylin may protect pancreatic cells in metabolic syndrome.

Shenhar-Tsarfaty S, Bruck T, Bennett ER, Bravman T, Aassayag EB, Waiskopf N, Rogowski O, Bornstein N, Berliner S, Soreq H - J. Cell. Mol. Med. (2011)

Bottom Line: However, the activity differences remained unexplained.We demonstrate that BChE interacts with amylin through its core domain and efficiently attenuates both amylin fibril and oligomer formation.Taken together, our results suggest that MetS-associated BChE increases could protect pancreatic β-cells in vivo by decreasing the formation of toxic amylin oligomers.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology and Internal Medicine, Tel Aviv Sourasky Medical Center, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.

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rhBChE and LOX, but not the BSP-U and BSP-K peptides, affect amylin fibril formation. Proteins and peptides were tested for their effect on amylin fibril formation kinetics by following ThT fluorescence (Excitation, 450 nm; emission, 485 nm) at room temperature for 25 hrs. The concentration of amylin in all experiments was 12.8 μM. The traces shown are representative of three separate experiments. (A) Amylin fibril formation in the absence (0 μM) and presence of 0.08 μM, 0.021 μM, 0.032 μM and 0.128 μM rhBChE. A dose-dependent attenuation of fibril formation is observed. (B) Amylin fibril formation in the absence (0 μM) and presence of 0.021 μM and 0.128 μM LOX. A less pronounced dose-dependent attenuation of fibril formation is observed. (C) Amylin fibril formation in the absence (0 μM) and presence of 0.032 μM, 0.128 μM and 0.512 μM BSP-U. No attenuation of fibril formation is observed. (D) Amylin fibril formation in the absence (0 μM) and presence of BSP-K (concentrations as in section C). As in the case of BSP-U, no attenuation of fibril formation is observed.
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fig04: rhBChE and LOX, but not the BSP-U and BSP-K peptides, affect amylin fibril formation. Proteins and peptides were tested for their effect on amylin fibril formation kinetics by following ThT fluorescence (Excitation, 450 nm; emission, 485 nm) at room temperature for 25 hrs. The concentration of amylin in all experiments was 12.8 μM. The traces shown are representative of three separate experiments. (A) Amylin fibril formation in the absence (0 μM) and presence of 0.08 μM, 0.021 μM, 0.032 μM and 0.128 μM rhBChE. A dose-dependent attenuation of fibril formation is observed. (B) Amylin fibril formation in the absence (0 μM) and presence of 0.021 μM and 0.128 μM LOX. A less pronounced dose-dependent attenuation of fibril formation is observed. (C) Amylin fibril formation in the absence (0 μM) and presence of 0.032 μM, 0.128 μM and 0.512 μM BSP-U. No attenuation of fibril formation is observed. (D) Amylin fibril formation in the absence (0 μM) and presence of BSP-K (concentrations as in section C). As in the case of BSP-U, no attenuation of fibril formation is observed.

Mentions: The similar amyloidogenic properties of amylin and Aβ[38] together with our previous findings that AChE-R can attenuate Aβ fibril formation in vivo [37] raised the possibility that rhBChE can similarly attenuate the aggregation of amylin. We therefore used the ThT fluorescence assay to measure the aggregation of amylin in the absence and presence of rhBChE. As shown in Figure 4A, amylin alone yielded a sigmoidal fluorescence time curve, consistent with the presence of lag, elongation and saturation phases in the fibril formation process, and in agreement with previous reports [10, 39]. Addition of rhBChE at increasing concentrations attenuated amylin fibril formation in a dose-dependent manner, reflected as gradually flattening curves with increasingly longer lag phases and lower saturation fluorescence levels, culminating in total prevention of fibril formation by 0.128 μM rhBChE. We next looked at the effect of LOX on amylin fibrillation. A possible role of LOX was proposed in both AD and T2DM: Similarly to BChE, the levels of LOX increase during the course of both these diseases [40, 41]. In addition, blockade of LOX expression can protect cortical neurons from Aβ-induced apoptosis [42] and knockout of LOX in mice can prevent cytokine-induced pancreatic β-cell dysfunction [43]. Intriguingly, as shown in Figure 4B, LOX also inhibited the fibrillation of amylin, albeit in a less potent manner than rhBChE, with a shorter lag phase and shallower elongation curve. The synthetic BSP-U and BSP-K peptides did not attenuate amylin fibril formation at any of the concentrations used (Fig. 4C and D), indicating that in contrast to its effect on Aβ, BChE attenuation of amylin oligomerization does not involve the C-terminal domain.


Butyrylcholinesterase interactions with amylin may protect pancreatic cells in metabolic syndrome.

Shenhar-Tsarfaty S, Bruck T, Bennett ER, Bravman T, Aassayag EB, Waiskopf N, Rogowski O, Bornstein N, Berliner S, Soreq H - J. Cell. Mol. Med. (2011)

rhBChE and LOX, but not the BSP-U and BSP-K peptides, affect amylin fibril formation. Proteins and peptides were tested for their effect on amylin fibril formation kinetics by following ThT fluorescence (Excitation, 450 nm; emission, 485 nm) at room temperature for 25 hrs. The concentration of amylin in all experiments was 12.8 μM. The traces shown are representative of three separate experiments. (A) Amylin fibril formation in the absence (0 μM) and presence of 0.08 μM, 0.021 μM, 0.032 μM and 0.128 μM rhBChE. A dose-dependent attenuation of fibril formation is observed. (B) Amylin fibril formation in the absence (0 μM) and presence of 0.021 μM and 0.128 μM LOX. A less pronounced dose-dependent attenuation of fibril formation is observed. (C) Amylin fibril formation in the absence (0 μM) and presence of 0.032 μM, 0.128 μM and 0.512 μM BSP-U. No attenuation of fibril formation is observed. (D) Amylin fibril formation in the absence (0 μM) and presence of BSP-K (concentrations as in section C). As in the case of BSP-U, no attenuation of fibril formation is observed.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4373355&req=5

fig04: rhBChE and LOX, but not the BSP-U and BSP-K peptides, affect amylin fibril formation. Proteins and peptides were tested for their effect on amylin fibril formation kinetics by following ThT fluorescence (Excitation, 450 nm; emission, 485 nm) at room temperature for 25 hrs. The concentration of amylin in all experiments was 12.8 μM. The traces shown are representative of three separate experiments. (A) Amylin fibril formation in the absence (0 μM) and presence of 0.08 μM, 0.021 μM, 0.032 μM and 0.128 μM rhBChE. A dose-dependent attenuation of fibril formation is observed. (B) Amylin fibril formation in the absence (0 μM) and presence of 0.021 μM and 0.128 μM LOX. A less pronounced dose-dependent attenuation of fibril formation is observed. (C) Amylin fibril formation in the absence (0 μM) and presence of 0.032 μM, 0.128 μM and 0.512 μM BSP-U. No attenuation of fibril formation is observed. (D) Amylin fibril formation in the absence (0 μM) and presence of BSP-K (concentrations as in section C). As in the case of BSP-U, no attenuation of fibril formation is observed.
Mentions: The similar amyloidogenic properties of amylin and Aβ[38] together with our previous findings that AChE-R can attenuate Aβ fibril formation in vivo [37] raised the possibility that rhBChE can similarly attenuate the aggregation of amylin. We therefore used the ThT fluorescence assay to measure the aggregation of amylin in the absence and presence of rhBChE. As shown in Figure 4A, amylin alone yielded a sigmoidal fluorescence time curve, consistent with the presence of lag, elongation and saturation phases in the fibril formation process, and in agreement with previous reports [10, 39]. Addition of rhBChE at increasing concentrations attenuated amylin fibril formation in a dose-dependent manner, reflected as gradually flattening curves with increasingly longer lag phases and lower saturation fluorescence levels, culminating in total prevention of fibril formation by 0.128 μM rhBChE. We next looked at the effect of LOX on amylin fibrillation. A possible role of LOX was proposed in both AD and T2DM: Similarly to BChE, the levels of LOX increase during the course of both these diseases [40, 41]. In addition, blockade of LOX expression can protect cortical neurons from Aβ-induced apoptosis [42] and knockout of LOX in mice can prevent cytokine-induced pancreatic β-cell dysfunction [43]. Intriguingly, as shown in Figure 4B, LOX also inhibited the fibrillation of amylin, albeit in a less potent manner than rhBChE, with a shorter lag phase and shallower elongation curve. The synthetic BSP-U and BSP-K peptides did not attenuate amylin fibril formation at any of the concentrations used (Fig. 4C and D), indicating that in contrast to its effect on Aβ, BChE attenuation of amylin oligomerization does not involve the C-terminal domain.

Bottom Line: However, the activity differences remained unexplained.We demonstrate that BChE interacts with amylin through its core domain and efficiently attenuates both amylin fibril and oligomer formation.Taken together, our results suggest that MetS-associated BChE increases could protect pancreatic β-cells in vivo by decreasing the formation of toxic amylin oligomers.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology and Internal Medicine, Tel Aviv Sourasky Medical Center, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.

Show MeSH
Related in: MedlinePlus