Myosin 51 heads

Myosin heads form cross-bridges between the actin and myosin filaments... 

Figure 14.15 Stmcture of the SI fragment of chicken myosin as a Richardson diagram (a) and a space-filling model (b). The two light chains are shown in magenta and yellow. The heavy chain is colored according to three proteolytic fragments produced by trypsin a 25-kDa N-terminal domain (green) a central 50-kDa fragment (red) divided by a cleft into a 50K upper and a 50K lower domain and a 20-kDa C-terminal domain (blue) that links the myosin head to the coiled-coil tail. The 50-kDa and 20-kDa domains both bind actin, while the 25-kDa domain binds ATP. [(b) Courtesy of 1. Rayment.]...

It seems likely, therefore, that as the bound phosphate molecule is released, the cleft starts to open and the myosin head binds to actin (Figure 14.17d). Release of ADP coincides with a conformational change that fully opens the myosin cleft, causing actin to be tightly bound, and moves the lever arm to the "down" position. Since the myosin head is now strongly bound to actin at one end and covalently linked to the myosin fibril at the other... 

Approximately 500 of the 820 amino acid residues of the myosin head are highly conserved between various species. One conserved region, located approximately at residues 170 to 214, constitutes part of the ATP-binding site. Whereas many ATP-binding proteins and enzymes employ a /3-sheet-a-helix-/3-sheet motif, this region of myosin forms a related a-f3-a structure, beginning with an Arg at (approximately) residue 192. The /3-sheet in this region of all myosins includes the amino acid sequence... 

The Coupling Mechanism ATP Hydrolysis Drives Conformation Changes in the Myosin Heads... 

FIGURE 17.23 The mechanism of skeletal muscle contraction. The free energy of ATP hydrolysis drives a conformational change in the myosin head, resulting in net movement of the myosin heads along the actin filament. Inset) A ribbon and space-filling representation of the actin—myosin interaction. (SI myosin image courtesy of Ivan Rayment and Hazel M. Holden, University of Wiseonsin, Madison.)... 

Molloy, J., Burns, J., Kendrick-Jones, J., et al., 1995. Movement and force produced by a single myosin head. Nature 378 209-213. 

The myosin head has long been shown to induce, even in low ionic strength buffers, polymerization of G-actin into decorated F-actin-S i filaments that exhibit the classical arrowhead structure (Miller et al., 1988 and older references therein). However, to date, the molecular mechanism of this polymerization process remains unknown. 

In an effort to understand how actin-actin interactions might be affected by the binding of the myosin head, and in order to gain more insight into the nature of the actin-myosin interface, we have investigated the nature of the kinetic actin-myosin intermediates involved in the process of S)-induced polymerization of G-actin. For this purpose, a variety of fluorescent probes (e.g., pyrene, NBD, AEDANS) have been covalently attached to the C-terminus of G-actin to probe the G-actin-S] interaction under conditions of tightest binding, i.e., in the absence of ATP. 

Chaussepied, P. Kasprzak, A.A. (1989). Isolation and characterization of the G-actin-myosin head complex. Namre 342,950-953. 

Manstein, D.J., Ruppel, K.M., Spudich, J.A. (1989). Expression and characterization of a functional myosin head fragment in Dictyostelium discoideum. Science 246, 656-658. 

First, in the striated muscles, the cross-sectional organization of filaments is highly ordered in a hexagonal pattern commensurate with the ratio of actin to myosin filaments and the distribution of active myosin heads, S-1 segments, helically every 60 degrees around the myosin filament. In smooth muscle, with perhaps 13 actin filaments per myosin filament, many actin filaments appear to be ranked in layers around myosin filaments. It is not known how the more distant actin filaments participate in contraction. 

Figure 49-4. Diagram of a myosin moiecuie showing the two intertwined a-heiices (fibrous portion), the giobuiar region or head (G),the iight chains (L), and the effects of proteoiytic cieavage by trypsin and papain. The giobuiar region (myosin head) contains an actin-binding site and an L chain-binding site and aiso attaches to the remainder of the myosin moiecuie.

When the sarcolemma is excited by a nerve impulse, the signal is transmitted into the T tubule system and a release channel in the nearby sarcoplasmic reticulum opens, releasing Ca + from the sarcoplasmic reticulum into the sarcoplasm. The concentration of Ca in the sarcoplasm rises rapidly to 10 mol/L. The Ca -binding sites on TpC in the thin filament are quickly occupied by Ca +. The TpC-4Ca + interacts with Tpl and TpT to alter their interaction with tropomyosin. Accordingly, tropomyosin moves out of the way or alters the conformation of F-actin so that the myosin head-ADP-P (Figure 49-6) can interact with F-actin to start the contraction cycle. 

Relaxation occurs when sarcoplasmic Ca falls below 10 mol/L owing to its resequestration into the sarcoplasmic reticulum by Ca ATPase. TpC.dCa thus loses its Ca. Consequently, troponin, via interaction with tropomyosin, inhibits further myosin head and F-actin interaction, and in the presence of ATP the myosin head detaches from the F-actin. 

A decrease in the concentration of ATP in the sarcoplasm (eg, by excessive usage during the cycle of con-traction-relaxation or by diminished formation, such as might occur in ischemia) has two major effects (1) The Ca ATPase (Ca + pump) in the sarcoplasmic reticulum ceases to maintain the low concentration of Ca + in the sarcoplasm. Thus, the Interaction of the myosin heads with F-actin is promoted. (2) The ATP-depen-dent detachment of myosin heads from F-actin cannot occur, and rigidity (contracmre) sets in. The condition of rigor mortis, following death, is an extension of these events. 

Muscle contraction is a delicate dynamic balance of the attachment and detachment of myosin heads to F-actin, subject to fine regulation via the nervous system. 

When smooth muscle myosin is bound to F-actin in the absence of other muscle proteins such as tropomyosin, there is no detectable ATPase activity. This absence of activity is quite unlike the situation described for striated muscle myosin and F-actin, which has abundant ATPase activity. Smooth muscle myosin contains fight chains that prevent the binding of the myosin head to F-actin they must be phosphorylated before they allow F-actin to activate myosin ATPase. The ATPase activity then attained hydrolyzes ATP about tenfold more slowly than the corresponding activity in skeletal muscle. The phosphate on the myosin fight chains may form a chelate with the Ca bound to the tropomyosin-TpC-actin complex, leading to an increased rate of formation of cross-bridges between the myosin heads and actin. The phosphorylation of fight chains initiates the attachment-detachment contraction cycle of smooth muscle. 

The hydrolysis of ATP is used to drive movement of the filaments. ATP binds to myosin heads and is hydrolyzed to ADP and P, by the ATPase activity of the actomyosin complex. 

Thick filaments. Each thick filament contains 200 to 300 myosin molecules. Each myosin molecule is made up of two identical subunits shaped like golf clubs two long shafts wound together with a myosin head, or crossbridge, on the end of each. These molecules are arranged so that the shafts are bundled together and oriented toward the center of the thick filament. The myosin heads project outward from either end of the thick filament (see Figure 11.1, panel a). 

Hiratsuka T (1986) Involvement of the 50-kDa peptide of myosin heads in the ATPase activity revealed by fluorescent modification with 4-fluoro-7-nitrobenzo-2-oxa-l,3-diazole. J Biol Chem 261 7294-7299... 

Labbe, J.R, Mornet, D., Roseau, G., and Kassab, R. (1982) Cross-linking of F-actin to skeletal muscle myosin subfragment 1 with bis(imido esters) Further evidence for the interaction of myosin-head heavy chain with an actin dimer. Biochemistry 21, 6897-6902. 

Walsh There s strong evidence that it is anchored to the actin filament through an N-terminal domain that Jim Stull s group has defined very precisely. The age-old question remains how does an anchored MLCK molecule gain access to an adequate number of myosin heads to account for the phosphorylation stoichiometry that can be achieved in muscle ... 

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