Myosin-binding protein C displaces tropomyosin to activate cardiac thin filaments and governs their speed by an independent mechanism.
Myosin-binding protein C (MyBP-C) is an accessory protein of striated muscle thick filaments and a modulator of cardiac muscle contraction. Defects in the cardiac isoform, cMyBP-C, cause heart disease. cMyBP-C includes 11 Ig- and fibronectin-like domains and a cMyBP-C-specific motif. In vitro studies show that in addition to binding to the thick filament via its C-terminal region, cMyBP-C can also interact with actin via its N-terminal domains, modulating thin filament motility. Structural observations of F-actin decorated with N-terminal fragments of cMyBP-C suggest that cMyBP-C binds to actin close to the low Ca(2+) binding site of tropomyosin. This suggests that cMyBP-C might modulate thin filament activity by interfering with tropomyosin regulatory movements on actin. To determine directly whether cMyBP-C binding affects tropomyosin position, we have used electron microscopy and in vitro motility assays to study the structural and functional effects of N-terminal fragments binding to thin filaments. 3D reconstructions suggest that under low Ca(2+) conditions, cMyBP-C displaces tropomyosin toward its high Ca(2+) position, and that this movement corresponds to thin filament activation in the motility assay. At high Ca(2+), cMyBP-C had little effect on tropomyosin position and caused slowing of thin filament sliding. Unexpectedly, a shorter N-terminal fragment did not displace tropomyosin or activate the thin filament at low Ca(2+) but slowed thin filament sliding as much as the larger fragments. These results suggest that cMyBP-C may both modulate thin filament activity, by physically displacing tropomyosin from its low Ca(2+) position on actin, and govern contractile speed by an independent molecular mechanism.
Department of Cell and Developmental Biology, University of Massachusetts Medical School, Worcester, MA 01655.
Ji Young
JY
Mun
Michael J
MJ
Previs
Hope Y
HY
Yu
James
J
Gulick
Larry S
LS
Tobacman
Samantha
S
Beck Previs
Jeffrey
J
Robbins
David M
DM
Warshaw
Roger
R
Craig
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002352:Carrier Proteins / Q000502:physiology*; D002645:Chickens; D008854:Microscopy, Electron; D009206:Myocardium / Q000378:metabolism*; D014335:Tropomyosin / Q000378:metabolism / Q000502:physiology*; D000099163:Myosin Binding Protein C
D002118:Calcium; D002352:Carrier Proteins; D014335:Tropomyosin; D000099163:Myosin Binding Protein C
muscle activation; muscle regulation