期刊
JOURNAL OF BIOLOGICAL CHEMISTRY
卷 288, 期 44, 页码 31952-31962出版社
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
DOI: 10.1074/jbc.M113.513291
关键词
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资金
- British Heart Foundation [PG/07/061/23277]
- Biotechnology and Biological Sciences Research Council [BB/I007423/1]
- Biotechnology and Biological Sciences Research Council
- BBSRC [BB/I007423/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/I007423/1] Funding Source: researchfish
It is unclear why mutations in the filament-forming tail of myosin heavy chain (MHC) cause hypertrophic or dilated cardiomyopathy as these mutations should not directly affect contraction. To investigate this, we first investigated the impact of five hypertrophic cardiomyopathy-causing (N1327K, E1356K, R1382W, E1555K, and R1768K) and one dilated cardiomyopathy-causing (R1500W) tail mutations on their ability to incorporate into muscle sarcomeres in vivo. We used adenoviral delivery to express full-length wild type or mutant enhanced GFP-MHC in isolated adult cardiomyocytes. Three mutations (N1327K, E1356K, and E1555K) reduced enhanced GFP-MHC incorporation into muscle sarcomeres, whereas the remainder had no effect. No mutations significantly affected contraction. Fluorescence recovery after photobleaching showed that fluorescence recovery for the mutation that incorporated least well (N1327K) was significantly faster than that of WT with half-times of 25.1 +/- 1.8 and 32.2 +/- 2.5 min (mean +/- S.E.), respectively. Next, we determined the effects of each mutation on the helical properties of wild type and seven mutant peptides (7, 11, or 15 heptads long) from the myosin tail by circular dichroism. R1382W and E1768K slightly increased the alpha-helical nature of peptides. The remaining mutations reduced alpha-helical content, with N1327K showing the greatest reduction. Only peptides containing residues 1301-1329 were highly alpha-helical suggesting that this region helps in initiation of coiled coil. These results suggest that small effects of mutations on helicity translate into a reduced ability to incorporate into sarcomeres, which may elicit compensatory hypertrophy.
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