Myosin synthesis

Okazaki K, Holtzer H (1966) Mogenesis, fusion, myosin synthesis and the mitotic cycle. Proc Natl Acad Sci USA 56 1484-1488... 

In a previous section we mentioned the significance of myosin filament structure. In nematodes two forms of myosin-II, myosin A and B, are required for proper filament stmcture (Epstein, 1988). The two forms of myosin are expressed at the proper time to allow for correct filament assembly. An accessory protein called paramyosin is also required for correct filament assembly. In vertebrate cardiac muscle, there are also two isoforms of myosin-II a-myosin and p-myosin. The proper ratio of these two proteins is of utmost importance for proper muscle activity. The incorrect synthesis of a- and P-myosins results in a severe cardiac disorder known as hypertrophic cardiomyopathy. Genetic transmission of the disease occurs in about 55% of families. The inherited condition is called familial hypertrophic cardiomyopathy (FHC), and this condition is a leading cause of sudden death in young athletes. 

The smdy of tissue protein breakdown in vivo is difficult, because amino acids released during intracellular breakdown of proteins can be extensively reutilized for protein synthesis within the cell, or the amino acids may be transported to other organs where they enter anabohc pathways. However, actin and myosin are methylated by a posttranslational reaction, forming d-methylliistidine. During intracellular breakdown of actin and myosin, 3-methylhistidine is released and excreted into the urine. The urinary output of the methylated amino acid provides a rehable index of the rate of myofibrillar protein breakdown in the musculature of human subjects. 

Because muscle fibers in males are thicker than those found in females, these muscles are larger and stronger, even without the benefit of resistance training. This enlargement is due to effects of testosterone, a sex hormone found primarily in males. Testosterone promotes the synthesis of actin and myosin filaments in muscle fibers. 

Figure 9.21 The creatine/phosphocreatine shuttle in spermatozoa. This shuttle may not be present in all sperm it will depend upon the distance between the mitochondria and the flagellum. Mitochondria are present in the midpiece just below the head. ATP is required for movement of the flagellum which enables the sperm to swim. Dynein ATPase is the specific motor ATPase, similar to myosin ATPase, that transfers energy from ATP to the flagellum. A deficiency of creatine may explain low sperm motility in some infertile men. CK - creatine kinase. Deficiences of enzymes in the pathway for synthesis of creatine are known to occur (see Appendix 8.3).

Figure 9.30 Flow diagram of the energy chain from food to essential processes in human life. The ATP utilised by the NayK ATPase maintains the ion distribution in nerves that is essential for electrical activity and, in addition, maintains neurotransmitter synthesis, both of which provide communication in the brain and hence consciousness, learning and behaviour (Chapter 14). ATP utilisation by myosin ATPase is essential for movement and physical activity. ATP utilisation by the flagellum of sperm is essential for reproduction and ATP utilisation for synthesis of macromolecules is essential for growth.

The long tail of myosin contains a high proportion of the amino acids leucine, isoleucine, aspartate and glutamate. These are released upon the degradation of myosin by intracellular proteases and peptidases and they provide nitrogen for the synthesis of glutamine. It is then stored in muscle and is a very important fuel for immune cells (Chapter 17). 

The idea of conformational coupling of ATP synthesis and electron transport is especially attractive when we recall that ATP is used in muscle to carry out mechanical work. Here we have the hydrolysis of ATP coupled to motion in the protein components of the muscle. It seems reasonable that ATP should be formed as a result of motion induced in the protein components of the ATPase. Support for this analogy has come from close structural similarities of the F, ATPase P subunits and of the active site of ATP cleavage in the muscle protein myosin (Chapter 19). 

Much is known about the steps in the biochemical reaction of ATP breakdown by myosin and how these relate to the production of force by the crossbridge. However, since it is no longer attached to the myosin thick filament, myosin SI cannot be an adequate model for a strained crossbridge. Thus data from muscle fibers (e.g., the dependence of phosphate affinity on strain) must also be considered. In this review we attempt to summarize the currently known structural data on myosin and produce a synthesis of this with the biochemical data. We start with an analysis of the polymorphism of the myosin crossbridge and relate this to the crossbridge cycle proposed by Lymn and Taylor (1971). 

Fig. 5. Regulation of myosin heavy chains by thyroid hormones during development and in adulthood. T, represses the synthesis of the fetal /3 myosin heavy chain mRNA while inducing the expression of the adult a isomyosin mRNA.

Airway smooth muscle cells isolated from canine tracheae and bronchi subjected to cyclic strain exhibit increased cell number and DNA synthesis in cell culture. The content of total cellular protein, especially contractile proteins including myosin, myosin light chain kinase, and desmin, was increased compared to cells cultured under static conditions. 

Rapid phosphorylation of the other detected phosphoproteins does occur but no definite roles have yet been ascribed to them. The 33 kDa protein may be the S6 ribosomal protein involved in the control of protein synthesis. The 57 kDa protein has been identified as the regulatory suhunit of the cyclic AMP-dependent protein kinase [44]. Of the other proteins the 76, 43 and 20 kDa may be connected with the microfilaments (43 kDa actin, 76 kDa myosin light chain kinase and 20 kDa myosin light chain) but this must be further investigated. These proteins may only play a permissive role in. steroidogenesis. The fact that the pattern of protein phosphorylation is very similar after stimulation of protein kinase C with phorbol esters supports this because the latter only marginally increase steroidogenesis [18]. 

Carnitine is synthesized from lysine and methionine by the pathway shown in Figure 14.2 (Vaz and Wanders, 2002). The synthesis of carnitine involves the stepwise methylation of a protein-incorporated lysine residue at the expense of methionine to yield a trimethyllysine residue. Free trimethyllysine is then released by proteolysis. It is not clear whether there is a specific precursor protein for carnitine synthesis, because trimethyllysine occurs in a number of proteins, including actin, calmodulin, cytochrome c, histones, and myosin. 

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