Enzyme skeletal muscle, release

Intravenous lipid emulsion particles are hydrolyzed in the bloodstream by the enzyme lipoprotein lipase to release free fatty acids and glycerol. Free fatty acids then are be taken up into adipose tissue for storage (triglycerides), oxidized to energy in various tissues (e.g., skeletal muscle), or recycled in the liver to make lipoproteins. 

Creatine kinase, creatine kinase myocardial band Creatine kinase (CK) enzymes are found in many isoforms, with varying concentrations depending on the type of tissue. Creatine kinase is a general term used to describe the nonspecific total release of all types of CK, including that found in skeletal muscle (MM), brain (BB) and heart (MB). CK MB is released into the blood from necrotic myocytes in response to infarction and is a useful laboratory test for diagnosing myocardial infarction. If the total CK is elevated, then the relative index (RI), or fraction of the total that is composed of CK MB, is calculated as follows RI = (CK MB/CK total) x 100. An RI greater than 2 is typically diagnostic of infarction. 

Both the G- and V-agents have the same physiological action on humans. They are potent inhibitors of the enzyme acetylcholinesterase (AChE), which is required for the function of many nerves and muscles in nearly every multicellular animal. Normally, AChE prevents the accumulation of acetylcholine after its release in the nervous system. Acetylcholine plays a vital role in stimulating voluntary muscles and nerve endings of the autonomic nervous system and many structures within the CNS. Thus, nerve agents that are cholinesterase inhibitors permit acetylcholine to accumulate at those sites, mimicking the effects of a massive release of acetylcholine. The major effects will be on skeletal muscles, parasympathetic end organs, and the CNS. 

The effect of Li+ upon the synthesis and release of acetylcholine in the brain is equivocal Li+ is reported to both inhibit and stimulate the synthesis of acetylcholine (reviewed by Wood et al. [162]). Li+ appears to have no effect on acetyl cholinesterase, the enzyme which catalyzes the hydrolysis of acetylcholine [163]. It has also been observed that the number of acetylcholine receptors in skeletal muscle is decreased by Li+ [164]. In the erythrocytes of patients on Li+, the concentration of choline is at least 10-fold higher than normal and the transport of choline is reduced [165] the effect of Li+ on choline transport in other cells is not known. A Li+-induced inhibition of either choline transport and/or the synthesis of acetylcholine could be responsible for the observed accumulation of choline in erythrocytes. This choline is probably derived from membrane phosphatidylcholine which is reportedly decreased in patients on Li+ [166],... 

The catecholamines can play an important role in the short-term regulation of plasma potassium levels. Stimulation of hepatic a-adrenoceptors will result in the release of potassium from the liver. In contrast, stimulation of (32-adrenoceptors, particularly in skeletal muscle, will lead to the uptake of potassium into this tissue. The (32-adrenoceptors are linked to the enzyme Na"", K+ adenosine triphosphatase (ATPase). Excessive stimulation of these (32-adrenoceptors may produce hypokalemia, which in turn can be a cause of cardiac arrhythmias. 

Activation of Gs or Gi proteins results in stimulation or inhibition, respectively, of adenylyl cyclase which catalyses the formation of cyclic adenosine monophosphate (cAMP) from ATP The cAMP binds to protein kinase A (PKA), which mediates the diverse cellular effects of cAMP by phosphorylating substrate enzymes, thereby increasing their activity. Among the responses mediated by cAMP are increases in contraction of cardiac and skeletal muscle and glycogenolysis in the liver by adrenaline (epinephrine). Because a single activated receptor can cause the conversion of up to 100 inactive Gs proteins to the active form, and each of these results in the synthesis of several hundred cAMP molecules, there is a very considerable signal amplification. For example, adrenaline concentrations as low as 10-10 M can stimulate the release of glucose sufficient to increase... 

As antioxidant peptides are rarely present in marine invertebrates, they must be released from the parent protein by hydrolysis with enzymes. Various enzymes have been used to release peptides from muscle proteins. To date, different muscle proteins have been extracted, hydrolysed, and their antioxidant activities studied, which is among all invertebrate muscles the most similar to vertebrate skeletal muscle. Various studies have been conducted to investigate the antioxidant properties of hydrolysates or bioactive peptides from marine invertebrate sources like oysters... 

Many diseases that cause tissue damage result in an increased release of intracellular enzymes into the plasma. The activities of many of these enzymes are routinely determined for diagnostic purposes in diseases of the heart, liver, skeletal muscle, and other tissues. The level of specific enzyme activity in the plasma frequently correlates with the extent of tissue damage. Thus, determining the degree of elevation of a particular enzyme activity in the plasma is often useful in evaluating the prognosis for the patient. 

The structure of an enzyme can also vary within a person, since different genes may encode enzymes that catalyse the same reaction. These enzymes are known as isozymes. Isozymes are often specific for different types of tissue. For example, lactate dehydrogenase (LDH) is produced in two forms, the M-type (muscle) and the H-type (heart). The M-type is predominates in tissue subject to anaerobic conditions, such as skeletal muscle and liver tissue, whereas the H-type predominates in tissue under aerobic conditions, such as the heart. Isozymes may be used as a diagnostic aid. For example, the presence of H-type LDH in the blood indicates a heart attack, since heart attacks cause the death of heart muscle with the subsequent release of H-type LDH into the circulatory system. 

An example of an enzyme which has different isoenzyme forms is lactate dehydrogenase (LDH) which catalyzes the reversible conversion of pyruvate into lactate in the presence of the coenzyme NADH (see above). LDH is a tetramer of two different types of subunits, called H and M, which have small differences in amino acid sequence. The two subunits can combine randomly with each other, forming five isoenzymes that have the compositions H4, H3M, H2M2, HM3 and M4. The five isoenzymes can be resolved electrophoretically (see Topic B8). M subunits predominate in skeletal muscle and liver, whereas H subunits predominate in the heart. H4 and H3M isoenzymes are found predominantly in the heart and red blood cells H2M2 is found predominantly in the brain and kidney while HM3 and M4 are found predominantly in the liver and skeletal muscle. Thus, the isoenzyme pattern is characteristic of a particular tissue, a factor which is of immense diagnostic importance in medicine. Myocardial infarction, infectious hepatitis and muscle diseases involve cell death of the affected tissue, with release of the cell contents into the blood. As LDH is a soluble, cytosolic protein it is readily released in these conditions. Under normal circumstances there is little LDH in the blood. Therefore the pattern of LDH isoenzymes in the blood is indicative of the tissue that released the isoenzymes and so can be used to diagnose a condition, such as a myocardial infarction, and to monitor the progress of treatment. 

Figure 2.7. The complex pathways and processes involved in fat catabolism in vertebrate tissues such as cardiac and skeletal muscles. FFAs arrive at the cell boundary either via VLDL or albumin-associated and enter the cell either by simple diffusion or through transporters. In the cytosol, FFAs are bound by FABPs, which increase the rate and amount of FFA that can be transferred to sites of utilization. Shorter chain FFAs are converted to acetylCoA in peroxisomes longer chain FFAs are directly transferred to mitochondria (via a complex system involving acylcarnitines) as long-chain acylCoA derivatives these enter the /6-oxidation spiral and are released as acetylCoA for entrance into the Krebs or citric acid cycle in the mitochondrial matrix. Fatty acid receptors (FARs) in the nucleus bind to fatty acid response elements (FAREs) and in turn regulate the production of enzymes in their own metabolism. (Modified from Veerkamp and Maatman, 1995.)...

The major substrate of phosphorylase b kinase is phosphorylase b which is phos-phorylated on a single serine residue at position 14, resulting in conversion to the more catalytically active form phosphorylase a [70], Phosphorylation of skeletal muscle phosphorylase also results in conversion of the Mr 200000 dimeric b form to the Mr 400000 tetrameric a form, whereas phosphorylation of the liver enzyme does not alter its dimeric structure [82]. Phosphorylase a is much less dependent than phosphorylase b upon the allosteric activator AMP [82], Since the activity of phosphorylase is rate-limiting for glycogen breakdown, its activation by phosphorylase b kinase results in enhanced glycogenolysis and glucose release from the liver. 

Type 1 NOS is a 168KDa protein found in the neurons which has been isolated and cloned [15]. This enzyme is responsible for the calcium-dependent release of NO from neurones and non-adrenergic, non-colinergic nerves and also from skeletal muscle [16]. The activity of type 1 and type III NOS enzymes is regulated by oestradiol which is a phenomenon observed during pregnancy [17]. 

Skeletal muscles also contribute enzymes to blood. Again, the cause may be poor perfusion, hypotliermia, or direct trauma to the muscles (crush injuries). Infection, inflammation (polymyositis), degenerative changes (dystrophies), drugs, and alcohol (alcoholic myopathy) wid cause enzyme leakage from myocytes. Enzyme release from muscles and other tissues also occurs as a result of anesthesia. 

Measurement of a wide range of serum enzymes in Duchenne dystrophy and comparison with the changes seen in other diseases has led Kleine (K7) to postulate that in this disease enzymes are released not only from skeletal muscle, but also from heart, liver, and possibly erythrocytes. 

Ghosh et al. [48] have isolated two isoforms of nSMase from rabbit skeletal muscle (92 and 53 kDa). Peptide mapping revealed important structural similarities, and the catalytic activities were also similar, except that the 53 kDa protein was Mg" -independent. These nSMases are located in the transverse tubules of the muscle cells, which may be related to the observation that sphingosine modulates calcium release from sarcoplasmic reticulum membranes [49]. Two Mn -and Mg" -dependent nSMases located in the microsomal membranes of seminiferous tubes of immature Wistar rats have been characterized [50] whose properties do not appear to differ significantly from other mammalian nSMases. Finally, we mention two other nSMases purified from eukaryotic natural sources, namely the Mg -dcpcridcril nSMase isolated from Saccharomyces cerevisiae [51], and that obtained from membrane fractions of intraeryfhrocytic Plasmodium falciparum, the malaria parasite [52]. The latter enzyme was activated by phosphatidylserine and other anionic phosphohpids, and was sensitive to scyphostatin, an inhibitor of mammalian nSMase (see below). 

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