Whether the two heads of skeletal muscle myosin work independently or cooperatively remains an open question in muscle biophysics. While individual myosin heads are sufficient for ATPase activity (Reisler (1980) J Mol Biol 138: 93-107) and force production (Harada et al. (1987) Nature 326: 805-808), it has also been reported that in situ, the two heads of a myosin molecule work cooperatively (Chaen et al. (1986) J Biol Chem 261(29): 13,632-13,636). To examine the role of cross-bridge cooperativity on isometric contraction and unloaded shortening we progressively inactivated myosin cross-bridges via titration with para-phenylenedimaleimide. The resting fiber ATPase was measured to provide an estimate of the fraction of active cross-bridges remaining during the titration. Isometric force and unloaded shortening velocity decline more rapidly than the resting ATPase as the titration proceeds. This is inconsistent with models for independent force generation and suggests cooperative action of myosin cross-bridges when muscle is isometrically contracting or shortening under zero load. However the degree of cooperativity depends on the type of muscle activity. While isometric force declines in a manner consistent with pair-wise cooperative action of myosin heads, unloaded shortening velocity declines more rapidly (greater cooperativity). Therefore, myosin cross-bridges in situ may be capable of at least two types of cooperative interactions, pair-wise cooperativity (when isometric) and another form of cooperativity that is sensitive to longer range interactions transmitted from other cross-bridges in the ensemble (during unloaded shortening).
Bibliographical noteFunding Information:
I would like to thank Cheryl Miller and Benjamin Trok for excellent technical assistance on this project. The project also benefited from the efforts of undergraduate honors students Leila Youakim and Chi Huynh who showed their dedication by mastering the ATPase assays. Dr. John Matta generously provided single fiber ATPase instruction and the fiber-mounting bracket. I would like to thank Dr. Mark Schoenberg who brought the project to my attention. This work was supported by funding from the Department of Physiology, the Dean of the University of Minnesota Medical School, and NIAMS (AR-43189).