The Structural Biology of Muscle: Spatial and Temporal Aspects

摘要

Understanding muscle contraction has resulted from the synergy of a number of approaches for which structure has provided an integrating framework. Nearly 60 years ago interference and phase contrast light microscopy established the sliding filament model of muscle contraction. A little later H.E. Huxley exploited electron microscopy to visualize the macromolecular architecture of the sarcomere: the thick (myosin) and thin (actin) filaments with connecting myosin cross-bridges. These observations allowed him to outline a structural basis for muscle contraction: a rowing-like progression of the myosin cross-bridges along the actin filament. X-ray fibre diffraction from insect flight muscle first demonstrated that the cross-bridges could indeed take up two configurations that might represent the ends of an active stroke. Later intense X-ray synchrotron radiation allowed the recording of the movements of the cross-bridges during a contraction with high precision. In 1993 Rayments's group ushered in a much more detailed understanding of myosin function by solving the structure of the myosin cross-bridge by X-ray crystallography. It showed that the cross-bridge consists of a large catalytic domain, often called the motor domain, containing the ATP binding site and the actin binding site. At the C-terminus of the motor domain is a long lever arm. The catalytic mechanism is similar to the G-proteins: the active site contains a P-loop and switch 1 and switch 2 elements. The lever arm was later found in two different conformations (so called pre-power-stroke and post-rigor) showing how switch 2 movement is coupled to a swing of the lever arm. Further cry stallographic studies coupled with high resolution em reconstructions of decorated actin (the rigor complex) showed how ATP binding sequesters switch 1 thereby opening the large cleft in the motor domain and breaking the strong binding to actin. Conversely, the strong binding to actin causes a movement of switch 1 with respect to the P-loop that destroys the nucleotide binding site, bringing about the release of ADP. There remains one unknown structure of seminal importance: the start the power stroke. During the cross-bridge cycle the cross-bridge in the pre-power stroke form loaded with ADP and phosphate rebinds to actin. The actin binding cleft must close without initially destroying the nucleotide binding site but in a way that enables phosphate release.