Journal article
Piperine, an alkaloid inhibiting the super-relaxed state of myosin, binds to the myosin regulatory light chain
Archives of biochemistry and biophysics, Vol.659, pp.75-84
12/01/2018
Handle:
https://hdl.handle.net/2376/114174
PMCID: PMC6699511
PMID: 30287237
Abstract
Piperine, an alkaloid from black pepper, was found to inhibit the super-relaxed state (SRX) of myosin in fast-twitch skeletal muscle fibers. In this work we report that the piperine molecule binds heavy meromyosin (HMM), whereas it does not interact with the regulatory light chain (RLC)-free subfragment-1 (S1) or with control proteins from the same muscle molecular machinery, G-actin and tropomyosin. To further narrow down the location of piperine binding, we studied interactions between piperine and a fragment of skeletal myosin consisting of the full-length RLC and a fragment of the heavy chain (HCF). The sequence of HCF was designed to bind RLC and to dimerize via formation of a stable coiled coil, thus producing a well-folded isolated fragment of the myosin neck. Both chains were co-expressed in Escherichia coli, the RLC/HCF complex was purified and tested for stability, composition and binding to piperine. RLC and HCF chains formed a stable heterotetrameric complex (RLC/HCF)2 which was found to bind piperine. The piperine molecule was also found to bind isolated RLC. Piperine binding to RLC in (RLC/HCF)2 altered the compactness of the complex, suggesting that the mechanism of SRX inhibition by piperine is based on changing conformation of the myosin.
•The SRX is crucial for maintaining the high economy of resting muscle.•Piperine destabilizes the SRX by a previously unknown mechanism.•We found that piperine binds to the regulatory light chain (RLC) of myosin.•Piperine destabilizes and reduces the compactness of the myosin neck.•This study facilitates the search for therapies for obesity and type 2 diabetes.
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Details
- Title
- Piperine, an alkaloid inhibiting the super-relaxed state of myosin, binds to the myosin regulatory light chain
- Creators
- Dmitri Tolkatchev - Voiland School of Chemical Engineering & Bioengineering, Washington State University, Pullman, WA, 99164-6515, USADaniel Elnatan - Department of Biochemistry/Biophysics, Univ. California, San Francisco, CA, 94158, USALeonardo Nogara - Department of Biochemistry/Biophysics, Univ. California, San Francisco, CA, 94158, USAThu Ly - Voiland School of Chemical Engineering & Bioengineering, Washington State University, Pullman, WA, 99164-6515, USANariman Naber - Department of Biochemistry/Biophysics, Univ. California, San Francisco, CA, 94158, USAKenny Haak - Voiland School of Chemical Engineering & Bioengineering, Washington State University, Pullman, WA, 99164-6515, USARyan Meech - Voiland School of Chemical Engineering & Bioengineering, Washington State University, Pullman, WA, 99164-6515, USARoger Cooke - Department of Biochemistry/Biophysics, Univ. California, San Francisco, CA, 94158, USAAlla S Kostyukova - Voiland School of Chemical Engineering & Bioengineering, Washington State University, Pullman, WA, 99164-6515, USA
- Publication Details
- Archives of biochemistry and biophysics, Vol.659, pp.75-84
- Academic Unit
- Chemical Engineering and Bioengineering, School of
- Publisher
- Elsevier Inc
- Identifiers
- 99900547841601842
- Language
- English
- Resource Type
- Journal article