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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, but not LOX, attenuates clearance of soluble amylin oligomers. Amylin was incubated in the absence and presence of proteins, the soluble phase was cross-linked using the PICUP method, and followed by SDS-PAGE separation and visualization by silver staining. (A) 51 μM amylin was incubated for 24 hrs in the absence and presence of 0.52 μM rhBChE, with aliquots removed at 0, 4, 8, 12 and 24 hrs. The gel shows the various oligomeric forms of amylin: the monomer is visible at ∼4 kD, and dimers, trimers, tetramers and pentamers are visible at the indicated positions. These forms are seen only in the samples containing rhBChE, visible at the top of the gel. The positions of the molecular mass standards are shown at the right. (B) 51 μM amylin was incubated for 8 hrs in the absence and presence of 0.52 μM LOX and aliquots were removed at 0, 4, 6 and 8 hrs. The gel shows the forms of amylin as in (A) LOX can be seen as a weak band at the top of the gel.
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fig05: rhBChE, but not LOX, attenuates clearance of soluble amylin oligomers. Amylin was incubated in the absence and presence of proteins, the soluble phase was cross-linked using the PICUP method, and followed by SDS-PAGE separation and visualization by silver staining. (A) 51 μM amylin was incubated for 24 hrs in the absence and presence of 0.52 μM rhBChE, with aliquots removed at 0, 4, 8, 12 and 24 hrs. The gel shows the various oligomeric forms of amylin: the monomer is visible at ∼4 kD, and dimers, trimers, tetramers and pentamers are visible at the indicated positions. These forms are seen only in the samples containing rhBChE, visible at the top of the gel. The positions of the molecular mass standards are shown at the right. (B) 51 μM amylin was incubated for 8 hrs in the absence and presence of 0.52 μM LOX and aliquots were removed at 0, 4, 6 and 8 hrs. The gel shows the forms of amylin as in (A) LOX can be seen as a weak band at the top of the gel.

Mentions: ThT fluorescence signals reflect the amount of amyloid fibrils formed [44] but do not define their exact oligomeric state. To determine this, we used the PICUP method to cross-link amylin monomers, then separated the resulting oligomeric complexes by SDS-PAGE and visualized them by silver staining. As can be seen in Figure 5A, at the initial time-point (only 3–5 min. after mixing the reaction components) amylin attained several soluble forms, from monomers up to hexamers, reflecting its highly amyloidogeic nature. These soluble forms were not visible, however, at later time-points. When amylin was incubated in the presence of rhBChE it showed similar oligomeric forms at the initial time-point, but these remained in the soluble fraction for at least 12 hrs. In comparison, when amylin was incubated with LOX (Fig. 5B) the oligomeric forms disappeared from the soluble fraction by 6 hrs. Thus, rhBChE prolonged the persistence of small amylin oligomers in solution, whereas LOX accelerated their precipitation out of solution.


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, but not LOX, attenuates clearance of soluble amylin oligomers. Amylin was incubated in the absence and presence of proteins, the soluble phase was cross-linked using the PICUP method, and followed by SDS-PAGE separation and visualization by silver staining. (A) 51 μM amylin was incubated for 24 hrs in the absence and presence of 0.52 μM rhBChE, with aliquots removed at 0, 4, 8, 12 and 24 hrs. The gel shows the various oligomeric forms of amylin: the monomer is visible at ∼4 kD, and dimers, trimers, tetramers and pentamers are visible at the indicated positions. These forms are seen only in the samples containing rhBChE, visible at the top of the gel. The positions of the molecular mass standards are shown at the right. (B) 51 μM amylin was incubated for 8 hrs in the absence and presence of 0.52 μM LOX and aliquots were removed at 0, 4, 6 and 8 hrs. The gel shows the forms of amylin as in (A) LOX can be seen as a weak band at the top of the gel.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4373355&req=5

fig05: rhBChE, but not LOX, attenuates clearance of soluble amylin oligomers. Amylin was incubated in the absence and presence of proteins, the soluble phase was cross-linked using the PICUP method, and followed by SDS-PAGE separation and visualization by silver staining. (A) 51 μM amylin was incubated for 24 hrs in the absence and presence of 0.52 μM rhBChE, with aliquots removed at 0, 4, 8, 12 and 24 hrs. The gel shows the various oligomeric forms of amylin: the monomer is visible at ∼4 kD, and dimers, trimers, tetramers and pentamers are visible at the indicated positions. These forms are seen only in the samples containing rhBChE, visible at the top of the gel. The positions of the molecular mass standards are shown at the right. (B) 51 μM amylin was incubated for 8 hrs in the absence and presence of 0.52 μM LOX and aliquots were removed at 0, 4, 6 and 8 hrs. The gel shows the forms of amylin as in (A) LOX can be seen as a weak band at the top of the gel.
Mentions: ThT fluorescence signals reflect the amount of amyloid fibrils formed [44] but do not define their exact oligomeric state. To determine this, we used the PICUP method to cross-link amylin monomers, then separated the resulting oligomeric complexes by SDS-PAGE and visualized them by silver staining. As can be seen in Figure 5A, at the initial time-point (only 3–5 min. after mixing the reaction components) amylin attained several soluble forms, from monomers up to hexamers, reflecting its highly amyloidogeic nature. These soluble forms were not visible, however, at later time-points. When amylin was incubated in the presence of rhBChE it showed similar oligomeric forms at the initial time-point, but these remained in the soluble fraction for at least 12 hrs. In comparison, when amylin was incubated with LOX (Fig. 5B) the oligomeric forms disappeared from the soluble fraction by 6 hrs. Thus, rhBChE prolonged the persistence of small amylin oligomers in solution, whereas LOX accelerated their precipitation out of solution.

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