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It is not possible to predict which infants are at risk for PBPP, and therefore amenable to preventive measures. Twenty-five per cent of affected infants will experience permanent impairment and injury. If recovery is incomplete by the end of the first month, referral to a multidisciplinary team is necessary. Further research into prediction, prevention and best mode of treatment needs to be done.

(RLC) in skeletal muscle has been proposed to act as a molecular memory of recent activation by increasing the rate of force development, ATPase activity, and isometric force at submaximal activation in fibers. It has been proposed that these effects stem from phosphorylation-induced movement of myosin heads away from the thick filament backbone. In this study, we examined the molecular effects of skeletal muscle myosin RLC phosphorylation using in vitro motility assays. We showed that, independently of the thick filament backbone, the velocity of skeletal muscle myosin is decreased upon phosphorylation due to an increase in the myosin duty cycle. Furthermore, we did not observe a phosphorylation-dependent shift in calcium sensitivity in the absence of the myosin thick filament. These data suggest that phosphorylation-induced movement of myosin heads away from the thick filament backbone explains only part of the observed phosphorylation-induced changes in myosin mechanics. Last, we showed that the duty cycle of skeletal muscle myosin is strain dependent, consistent with the notion that strain slows the rate of ADP release in striated muscle.in vitro motility assay; mechanics ALL KNOWN MUSCLE SYSTEMS share the characteristic that the regulation of actomyosin interaction is mediated by the gated release of calcium. The specific mechanism by which calcium regulates this interaction depends on the muscle type. In molluscan muscle, calcium binds directly to the myosin essential light chain (ELC) that, together with the regulatory light chain (RLC) and part of the myosin heavy chain (MHC), constitutes the myosin regulatory domain that switches on the myosin motor in response to calcium binding (86). In vertebrate cardiac and skeletal muscle, calcium binds to troponin C, causing a shift in the position of tropomyosin along actin and allowing myosin cross bridges to bind strongly to the thin filament and shorten the sarcomere (for review, see Ref. 22). Smooth muscle has yet another pathway for activation in which calcium binding to calmodulin activates myosin light chain kinase (MLCK), phosphorylating the RLC and activating smooth muscle contraction (for review, see Ref. 70).It was shown by Perrie et al. (50) that vertebrate striated muscle RLC could also be phosphorylated by activated MLCK, and although this interaction is not necessary for activation of contraction, it appears to modulate skeletal and cardiac muscle contractility (40). Phosphorylation of the RLC in striated muscle fibers has been shown to enhance both the magnitude (13,52,66,74,79) and rate of tension development (42, 73) as well as to increase the ATPase rate (79) at submaximal calcium levels (for review, see Ref. 71). Furthermore, it has been shown that myosin phosphorylation can cause potentiation of posttetanic twitch (39, 40) and the rate of cross-bridge attachment (14,42,48,73,79). These effects have also been shown to be removed by knocking out skeletal muscle MLCK (89). The extent of RLC phosphorylation correlates with the frequency of act...

The glutamic acid to lysine mutation at the 22nd amino acid residue (E22K) in the human cardiac myosin regulatory light chain (RLC) gene causes familial hypertrophic cardiomyopathy (FHC) with a phenotype of midventricular obstruction and septal hypertrophy. Our recent histopathology results have shown that the hearts of transgenic E22K mice (Tg-E22K) resemble those of human patients, demonstrating enlarged interventricular septa and papillary muscles. In this study, we show no effect of the E22K mutation on the kinetics of mutated myosin in its ATP-powered interaction with fluorescently labeled single actin filaments compared to nontransgenic or transgenic wild-type (Tg-WT) control mice. Likewise, no change in cross-bridge dissociation rates (g(app)) was observed in freshly skinned papillary muscle fibers. In contrast, maximal force and ATPase were decreased approximately 20% in Tg-E22K skinned papillary muscle fibers and intracellular [Ca2+] and force transients were significantly decreased in intact papillary muscle fibers from Tg-E22K compared to Tg-WT mice. Moreover, energy metabolism measured in isolated working Tg-E22K mouse hearts perfused under conditions of physiologically relevant levels of metabolic demand was similar in Tg-E22K and control hearts before and after 20 min of no-flow ischemia. Our results suggest that the pathological response observed in the E22K myocardium might be triggered by mutation induced changes in the properties of the RLC Ca2+-Mg2+ site, the state of the Ca2+/Mg2+ occupancy and consequently the Ca2+ buffering ability of the RLC. By decreasing the affinity of the RLC for Ca2+, the E22K mutation most likely promotes a Mg2+-saturated RLC producing less force and ATPase than the Ca2+-saturated RLC of WT fibers. Decreased Ca2+ binding may also lead to faster Ca2+ dissociation kinetics in Tg-E22K intact fibers resulting in decreased duration and amplitude of [Ca2+] and force transients. These changes when placed in vivo would result in higher workloads and consequently cardiac hypertrophy.

The myosin regulatory light chain (RLC) wraps around the alpha helical neck region of myosin. This neck region has been proposed to act as a lever arm, amplifying small conformational changes in the myosin head to generate motion. The RLC serves an important structural role, supporting the myosin neck region and a modulatory role, tuning the kinetics of the actin myosin interaction. Given the importance of the RLC, it is not surprising that mutations of the RLC can lead to familial hypertrophic cardiomyopathy (FHC), the leading cause of sudden cardiac death in people under 30. Population studies identified two FHC mutations located near the cationic binding site of the RLC, R58Q and N47K. Although these mutations are close in sequence, they differ in clinical presentation and prognosis with R58Q showing a more severe phenotype. We examined the molecular based changes in myosin that are responsible for the disease phenotype by purifying myosin from transgenic mouse hearts expressing mutant myosins and examining actin filament sliding using the in vitro motility assay. We found that both R58Q and N47K showed reductions in force compared to the wild type that could result in compensatory hypertrophy. Furthermore, we observed a higher ATPase rate and an increased activation at submaximal calcium levels for the R58Q myosin that could lead to decreased efficiency and incomplete cardiac relaxation, potentially explaining the more severe phenotype for the R58Q mutation.