Force phosphorylation

The thylakoid membrane is asymmetrically organized, or sided, like the mitochondrial membrane. It also shares the property of being a barrier to the passive diffusion of H ions. Photosynthetic electron transport thus establishes an electrochemical gradient, or proton-motive force, across the thylakoid membrane with the interior, or lumen, side accumulating H ions relative to the stroma of the chloroplast. Like oxidative phosphorylation, the mechanism of photophosphorylation is chemiosmotic. 

Hafher, R.P., Brown, G.C.. Brand, M.D. (1990). Analysis of the control of respiration rate, phosphorylation rate, proton leak rate and proton motive force in isolated mitochondria using the top-down approach of metabolic control theory. Eur. J. Biochem. 188,313-319. 

Although in in vivo circumstances an intracellular free calcium increase apparently acts as the primary modulator of contraction, it can be bypassed in highly permeabilized smooth muscle preparations where the active subunit of MLCK can be introduced to phosphorylate myosin and induce contraction. The MLCK catalyzed phosphorylation of serine-19 is seen as the necessary event in the activation of smooth muscle myosin to form crossbridges. Thus, the rising phase of force during an isometric smooth muscle contraction follows an increase in the degree of phosphorylation of myosin, and that in turn follows the transient rise of (a) cytosolic free Ca, (b) Ca-calmodulin complexes, and (c) the active form of MLCK. The regulation of the intracellular calcium is discussed below. The dynam-... 

One should note overall, that while some of these suggested mechanisms may in the future prove to have a role in the control of smooth muscle contraction, in chemically skinned preparations maximum force development follows activation by the MLCK active subunit in extremely low Ca " ion concentrations. The conclusion can hardly be avoided that phosphorylation alone is sufficient for activation, and if another mechanism is involved, it is not necessary for the initial genesis of force. If such mechanisms are operative, then they might be expected to run in parallel or consequent to myosin phosphorylation. A possible example of this category of effect is that a GTP-dependent process (G-protein) shifts the force vs. Ca ion concentration relationship to lower Ca ion concentrations. This kind of mechanism calls attention to the divergence of signals along the intracellular control pathways. 

Figure 8. For any set of conditions, the greatest velocity that a muscle can shorten is attained when the total force opposing shortening is zero. Empirically, the maximum velocity of shortening increases with the degree of phosphorylation of myosin. This is seen as the straight line in the velocity-phosphorylation plane. The maximum force that a smooth muscle can develop is not increased by phosphorylation beyond about 25% phosphorylation. It seems therefore that past a point, phosphorylation regulates the rate at which work is being done rather than the force that can be developed. The force a muscle can develop if 25% myosin is phosphorylated is maximal and saturated however, the rate of doing work is not saturated and continues to increase with further phosphorylation.

This potential, or protonmotive force as it is also called, in turn drives a number of energy-requiring functions which include the synthesis of ATP, the coupling of oxidative processes to phosphorylation, a metabohc sequence called oxidative phosphorylation and the transport and concentration in the cell of metabolites such as sugars and amino acids. This, in a few simple words, is the basis of the chemiosmotic theory linking metabolism to energy-requiring processes. 

Certain chemical substances have been known for many years to uncouple oxidation firm phosphorylation and to inhibit active transport, and for this reason they are named imcoupling agerrts. They are beheved to act by rendering the membrane permeable to protons hence short-circuiting the potential gradient or protonmotive force. 

Allelopathic inhibition of mineral uptake results from alteration of cellular membrane functions in plant roots. Evidence that allelochemicals alter mineral absorption comes from studies showing changes in mineral concentration in plants that were grown in association with other plants, with debris from other plants, with leachates from other plants, or with specific allelochemicals. More conclusive experiments have shown that specific allelochemicals (phenolic acids and flavonoids) inhibit mineral absorption by excised plant roots. The physiological mechanism of action of these allelochemicals involves the disruption of normal membrane functions in plant cells. These allelochemicals can depolarize the electrical potential difference across membranes, a primary driving force for active absorption of mineral ions. Allelochemicals can also decrease the ATP content of cells by inhibiting electron transport and oxidative phosphorylation, which are two functions of mitochondrial membranes. In addition, allelochemicals can alter the permeability of membranes to mineral ions. Thus, lipophilic allelochemicals can alter mineral absorption by several mechanisms as the chemicals partition into or move through cellular membranes. Which mechanism predominates may depend upon the particular allelochemical, its concentration, and environmental conditions (especially pH). 

The effect of stimulation of cardiac adrenoceptors is even more leisurely because several more steps follow activation of the Gs protein by the p-adrenoceptor. For example, to increase the force of cardiac contraction, we have (1) activation of adenylate cyclase by Gas-GTP, (2) formation of cAMP, (3) activation of protein kinase A by the cAMP, then (4) phosphorylation of the calcium channel protein by the kinase. As a result, it takes about 5 to 6 sec from the time the receptors are... 

Ca2+ entry, Ca2+-uptake into the SR by SERCA, Ca2+ extrusion from the cell and dephosphorylation of the myosin light chains. The t ype 1 phosphatase, myosin light chain phosphatase (MLCP) dephosphorylates myosin. As with MLCK its activity is physiologically regulated, e.g. its activity is decreased following phosphorylation via Rho associated kinase (Somlyo Somlyo 2000). In the uterus we have found a small but significant reduction of force, but not Ca2+ when Rho-associated kinase is inhibited (Kupittayanant et al 2001b). 

Hellstrand That is what I am getting at. There are a lot of phase shifts in this system. One observation we have made is that under hypoxia we see a decrease in amplitude but an increase in frequency of the waves. We are trying to model a case where this would account for reduction of force simply on the basis of non-linearity of the [Ca2+] versus myosin phosphorylation versus force reactions. It seems intuitively that this could explain why there can be a reduction in force although there is no reduction in the overall level of global Ca2+. Is amplitude modulation something that people have seen ... 

Wier It seems reasonable to think that these waves may serve to spike up the phosphorylation of myosin light chain. In combination with the Ca2+ sensitizing mechanisms, this force can be maintained. 

In order to synthesize biologically relevant phosphonylimidazole 73, bromoimidazole 72 was derived from radical-initiated bromination of methyl l-p-methoxybenzyl-2-thiomethyl-5-imidazolylcarboxylate (71) [56]. The thiomethyl group served to block the C(2) position, which would otherwise undergo preferential halogenation under these conditions. As expected, a variety of Arbusov-Michaelis reaction conditions failed even under forcing conditions. On the other hand, Pd-catalyzed phosphorylation of 72 with diethyl phosphite led to methyl-4-diethylphosphonyl-l-p-methoxybenzyl-2-thiomethyl-5-imidazolylcarboxylate (73). After further manipulations, the desired phosphonic acid-linked aminoimidazoles, which resembled intermediates formed during purine biosynthesis, were accessed. 

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