Contractile proteins from smooth muscle

The principal molecular constituent of thin filaments is actin. Actin has been highly conserved during the course of evolution and is present in all eukaryotes, including single-celled organisms such as yeasts. Actin was first extracted and purified from skeletal muscle, where it forms the thin filaments of sarcomeres. It also is the main contractile protein of smooth muscle. Refined techniques for the detection of small amounts of actin (e.g., immunofluorescence microscopy, gel electrophoresis, and EM cytochemistry) subsequently confirmed the presence of actin in a great variety of nonmuscle cells. Muscle and nonmuscle actins are encoded by different genes and are isoforms. 

A role for MAPK in the contractile responsiveness of smooth muscle may result from either of two possible mechanisms, both involving caldesmon phosphorylation. First, caldesmon phosphorylation by MAPK may lead directly to an alteration of actomyosin activity. Caldesmon may exert this effect alone, or in concert with other myofibrillar proteins such as cal-ponin. Second, phosphorylation of caldesmon may alter the dynamics of actin filament organization within the cell. Caldesmon phosphorylation may result in alterations of the cellular cytoskeleton that must occur during prolonged contractions. 

The smooth muscle cell does not respond in an all-or-none manner, but instead its contractile state is a variable compromise between diverse regulatory influences. While a vertebrate skeletal muscle fiber is at complete rest unless activated by a motor nerve, regulation of the contractile activity of a smooth muscle cell is more complex. First, the smooth muscle cell typically receives input from many different kinds of nerve fibers. The various cell membrane receptors in turn activate different intracellular signal-transduction pathways which may affect (a) membrane channels, and hence, electrical activity (b) calcium storage or release or (c) the proteins of the contractile machinery. While each have their own biochemically specific ways, the actual mechanisms are for the most part known only in outline. 

If MLCK activates contraction by increasing myosin phosphorylation, then an increase in the activity of myosin light chain phosphatase, MLCP, by decreasing the fraction of myosin which is phosphorylated, should lead to relaxation from the active (contractile) state. Cyclic adenosine monophosphate (AMP) is a strong inhibitor of smooth muscle contraction and it has been suggested that activation of MLCP could result from its phosphorylation via cAMP activated protein kinase (see Figure 5). 

Smooth muscle differs from skeletal muscle in various ways. Smooth muscles—which are found, for example, in blood vessel walls and in the walls of the intestines—do not contain any muscle fibers. In smooth-muscle cells, which are usually spindle-shaped, the contractile proteins are arranged in a less regular pattern than in striated muscle. Contraction in this type of muscle is usually not stimulated by nerve impulses, but occurs in a largely spontaneous way. Ca (in the form of Ca -calmodulin see p.386) also activates contraction in smooth muscle in this case, however, it does not affect troponin, but activates a protein kinase that phosphorylates the light chains in myosin and thereby increases myosin s ATPase activity. Hormones such as epinephrine and angiotensin II (see p. 330) are able to influence vascular tonicity in this way, for example. 

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. 

Most likely, proteins of the actomyosin type also exist in other smooth muscles. litiegg and Strassner (1963), for instance, have isolated from arterial walls a protein which is soluble at high ionic strengths and which exhibits ATPase activity. More detailed ijiformation about this material is not yet available, and therefore a comparison with the other contractile proteins is not possible at the present time. 

The contraction of ascidian smooth muscle was found to be regulated through the troponin-tropomyosin system. But the action of troponin components was different from that of troponin of vertebrate striated muscles (Endo and Obinata, 1981). In this system, the inhibitory action of troponin I (MW 24,000) is less remarkable compared with vertebrate skeletal troponin I, and troponin C (MW 18,000) does not neutralize the inhibition by troponin I. But upon further addition of troponin T (MW 33,000) in the concomitant presence of all three components and tropomyosin, the contractile interaction of myosin and actin is activated. In this case, the action of troponin T has some similarity with that of the above-mentioned cardiac troponin T hybridized with skeletal troponin C-I. Since actomyosin, without these regulatory proteins, is inhibited regardless of Ca concentration, Ca " and troponin-tropomyosin are activators for contraction of actomyosin in ascidian smooth muscle. In this respect, the type of Ca + regulation of ascidian smooth muscle is the same as that for vertebrate smooth muscles which do not contain troponin (Ebashi, 1980). 

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