Converting the Highly Amyloidogenic Human Calcitonin into a Powerful Fibril Inhibitor by Three-dimensional Structure Homology with a Non-amyloidogenic Analogue

被引:34
作者
Andreotti, Giuseppina [1 ]
Vitale, Rosa Maria [1 ]
Avidan-Shpalter, Carmit [2 ]
Amodeo, Pietro [1 ]
Gazit, Ehud [2 ]
Motta, Andrea [1 ]
机构
[1] CNR, Ist Chim Biomol, Comprensorio Olivetti, I-80078 Pozzuoli, Naples, Italy
[2] Tel Aviv Univ, Dept Mol Microbiol & Biotechnol, George S Wise Fac Life Sci, IL-69978 Tel Aviv, Israel
关键词
SALMON-CALCITONIN; SECONDARY STRUCTURE; ALPHA-HELIX; PROTEIN AGGREGATION; DYNAMIC PROPERTIES; NMR-SPECTROSCOPY; BETA PEPTIDE; FIBRILLOGENESIS; OLIGOMERIZATION; DETERMINANTS;
D O I
10.1074/jbc.M110.182014
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Irreversible aggregation limits bioavailability and therapeutic activity of protein-based drugs. Here we show that an aggregation-resistant mutant can be engineered by structural homology with a non-amyloidogenic analogue and that the aggregation-resistant variant may act as an inhibitor. This strategy has successfully been applied to the amyloidogenic human calcitonin (hCT). Including only five residues from the non-amyloidogenic salmon calcitonin (sCT), we obtained a variant, polar human calcitonin (phCT), whose solution structure was shown by CD, NMR, and calculations to be practically identical to that of sCT. phCT was also observed to be a potent amyloidogenesis inhibitor of hCT when mixed with it in a 1:1 ratio. Fibrillation studies of phCT and the phCT-hCT mixture mimicked the sCT behavior in the kinetics and shapes of the fibrils with a dramatic reduction with respect to hCT. Finally, the effect of phCT alone and of the mixture on the intracellular cAMP level in T47D cells confirmed for the mutant and the mixture their calcitonin-like activity, exhibiting stimulation effects identical to those of sCT, the current therapeutic form. The strategy followed appears to be suitable to develop new forms of hCT with a striking reduction of aggregation and improved activity. Finally, the inhibitory properties of the aggregation-resistant analogue, if confirmed for other amyloidogenic peptides, may favor a new strategy for controlling fibril formation in a variety of human diseases.
引用
收藏
页码:2707 / 2718
页数:12
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