Creatine, phosphorylation

Urea is formed in the body from ammonia that comes from deamination of amino acids it permeates throngh the entire body except the brain. Excreted urea can be used as a nitrogen source by microbes, plants, and animals. Creatinine is the nitrogenous waste product of muscle creatine phosphorylated creatine (also called phosphageri) is an energy source alternative to ATP (see Section 3.9). Diadrast is the commercial name for iodopyracet, an iodinated dye used to determine kidney function it has the property that a very high proportion is filtered and excreted from the kidney in a very short time. Hippurate is a salt of hippuric acid, and is used as a test of liver function. 

Hypoxia exerts a reciprocal control on transcription of glycolytic (increase) and mitochondrial (decrease) enzymes in myotubes (Webster et al. 1990). The effects on glycolytic enzymes may be mediated by the oxygen sensitive transcription factor HIE (hypoxia-inducible factor). In the immorta-Used mouse skeletal muscle cell line (C2C12), when used at low concentrations, azide inhibited more than 70 % of cytochrome oxidase activity without changes in bioenergetics (either lactate production or creatine phosphorylation) or mRNA for mitochondrial enzymes (Leary etal. 1998). 

A good example of an affinity label for creatine kinase has been presented (35). This enzyme catalyzes the reversible transfer of a phosphoryl group from adenosine triphosphate [56-65-5] (17) to creatine [57-00-1] (18), leading to adenosine diphosphate [7584-99-8] (19) and phosphocreatine [67-07-2]... 

An example of a random, single-displacement mechanism is seen in the enzyme creatine kinase, a phosphoryl-transfer enzyme that uses ATP as a phosphoryl... 

The lower panel shows the decreasing concentration of ATP, to about 60% of resting levels, and the simultaneous equimolar increase in IMP. The fall in ATP started when most of the PCr store was utilized, resulting in a decreased rate of ADP phosphorylation via the creatine kinase reaction. The resultant accumulation of ADP stimulates adenylate kinase activity and subsequently IMP is formed via the AMP deaminase reaction ... 

Creatine supplementation has been shown to increase the rate of PCr resynthesis in the recovery period after ischemic exercise (Greenhaff et al., 1993a). This could be attributed to an acceleration of oxidative phosphorylation by increased free creatine content available to the mitochondrial fraction of the creatine kinase enzyme, as previously suggested (Bessman and Fonyo, 1966). 

Figure 12-14. The creatine phosphate shuttle of heart and skeletal muscle. The shuttle allows rapid transport of high-energy phosphate from the mitochondrial matrix into the cytosol. CKg, creatine kinase concerned with large requirements for ATP, eg, muscular contraction CIC, creatine kinase for maintaining equilibrium between creatine and creatine phosphate and ATP/ADP CKg, creatine kinase coupling glycolysis to creatine phosphate synthesis CK, , mitochondrial creatine kinase mediating creatine phosphate production from ATP formed in oxidative phosphorylation P, pore protein in outer mitochondrial membrane.

The ATP required as the constant energy source for the contraction-relaxation cycle of muscle can be generated (1) by glycolysis, using blood glucose or muscle glycogen, (2) by oxidative phosphorylation, (3) from creatine... 

Creatine phosphate is formed from ATP and creatine (Figure 49-16) at times when the muscle is relaxed and demands for ATP are not so great. The enzyme catalyzing the phosphorylation of creatine is creatine kinase (CK), a muscle-specific enzyme with clinical utility in the detection of acute or chronic diseases of muscle. 

A Sprinter Uses Creatine Phosphate Anaerobic Glycolysis to Make ATP, Whereas a Marathon Runner Uses Oxidative Phosphorylation... 

CK catalyzes the reversible phosphorylation of creatine in the presence of ATP and magnesium. When creatine phosphate is the substrate, the resulting creatine can be measured as the ninhydrin fluorescent compound, as in the continuous flow Auto Analyzer method. Kinetic methods based on coupled enzymatic reactions are also popular. Tanzer and Gilvarg (40) developed a kinetic method using the two exogenous enzymes pyruvate kinase and lactate dehydrogenase to measure the CK rate by following the oxidation of NADH. In this procedure the main reaction is run in a less favorable direction. 

Explain how creatine phosphate, oxidative phosphorylation, and glycolysis provide energy for skeletal muscle contraction... 

During the recovery period from exercise, ATP (newly produced by way of oxidative phosphorylation) is needed to replace the creatine phosphate reserves — a process that may be completed within a few minutes. Next, the lactic acid produced during glycolysis must be metabolized. In the muscle, lactic acid is converted into pyruvic acid, some of which is then used as a substrate in the oxidative phosphorylation pathway to produce ATP. The remainder of the pyruvic acid is converted into glucose in the liver that is then stored in the form of glycogen in the liver and skeletal muscles. These later metabolic processes require several hours for completion. 

CNDO/2 Theoretical calculations have been used to predict the favoured conformations of creatine and creatine derivatives, including phosphocreatine. These calculations predict that phosphocreatine may adopt conformation (35) to avoid unnecessary steric and electrostatic repulsions.110 The possibility also exists that creatine kinase phosphorylates creatine stereospecifically to form the favoured conformation (35) at the active site of the enzyme. When chicks are fed a diet containing cyclocreatine (l-carboxymethyl-2-iminoimidazole), the phosphorylated... 

During periods of recovery following exercise, creatine phosphate is regenerated at the expense of ATP synthesized from mitochondrial oxidative phosphorylation energy currency is paid into a reserve account, or reservoir, for the next period of sustained exercise. 

The synthesis of creatine. In the kidney, guanidinoace-tate is produced from arginine and glycine, then released into the blood to be taken up by the liver and methylated to form creatine (Figure 8.20(a)). The creatine is, in turn, taken up by the muscle where it is phosphorylated to produce phosphocreatine, which can maintain the ATP level, especially in explosive exercise. Creatine and phosphocreatine are converted in muscle to creatinine, which is important in clinical practice (Figure 8.20(b)) (Box 8.3). 

Figure 8.20 (a) The synthesis of phosphocreatine. The compound guanidinoacetate is formed from arginine and glycine in the kidney and is then transported to the liver where it is methylated addition of CHj (see Chapter 15) to form creatine (see Appendix 8.4 for details). Creatine is taken up by tissues/ organs/cells and phosphorylated to form phosphocreatine, particularly in muscle, (b) Conversion of phosphocreatine and creatine to creatinine in muscle. Creatinine is gradually formed and then released into blood and excreted in urine. 

The ATP, which is transported out of the mitochondrion, immediately phosphorylates creatine, catalysed by the creatine kinase in the intermembrane space. 

At this site, creatine kinase catalyses the phosphorylation of ADP by phosphocreatine with the production of ATP which, in turn, is used by the energy-requiring process that is, the cross-bridge cycle (Chapter 13). 

Creatine (N-methylguanidoacetic acid) and its phosphorylated form creatine phosphate (a guanidophosphate) serve as an ATP buffer in muscle metabolism. In creatine phosphate, the phosphate residue is at a similarly high chemical potential as in ATP and is therefore easily transferred to ADP. Conversely, when there is an excess of ATP, creatine phosphate can arise from ATP and creatine. Both processes are catalyzed by creatine kinase [5]. 

In resting muscle, creatine phosphate forms due to the high level of ATP. If there is a risk of a severe drop in the ATP level during contraction, the level can be maintained for a short time by synthesis of ATP from creatine phosphate and ADP. In a nonenzymatic reaction [6], small amounts of creatine and creatine phosphate cyclize constantly to form creatinine, which can no longer be phosphorylated and is therefore excreted with the urine (see p. 324). 

In the course of studying the mechanism of action of creatine kinase from rabbit skeletal muscle (M.M isoenzyme), Kenyon and coworkers (4,90) have been involved in the design of specific irreversible inhibitors that are active-site-directed (affinity labels). Creatine kinase catalyzes the reversible transfer of a phosphoryl group ( the elements of "POi") from ATP to creatine, as shown in the following reaction ... 

Creatine (Cr) plays an important role in energy transmission and storage in cells and tissues with high energy demands. Tissues like the brain, retina, spermatozoa and cardiac and skeletal muscle contain the enzyme Cr kinase, which catalyses the interconversion of Cr and its phosphorylated analogue, phosphocreatine. The dephosphorylation of phosphocreatine yields energy, as ADP is simultaneously converted into ATP. 

Gellerich, F.N. Laterveer, F.D. Korzeniewski, B. Zierz, S. Nicolay, K. Dextran strongly increases the Michaelis constants of oxidative phosphorylation and of mitochondrial creatine kinase in heart mitochondria. Eur. J. Biochem., 254, 172-180 (1998)... 

Phosphocreatine (Fig. 13-5), also called creatine phosphate, serves as a ready source of phosphoryl groups for the quick synthesis of ATP from ADP. The phosphocreatine (PCr) concentration in skeletal muscle is approximately 30 nra, nearly ten times the concentration of ATP, and in other tissues such as smooth muscle, brain, and kidney [PCr] is 5 to 10 mM. The enzyme creatine kinase catalyzes the reversible reaction... 

When a sudden demand for energy depletes ATP, the PCr reservoir is used to replenish ATP at a rate considerably faster than ATP can be synthesized by catabolic pathways. When the demand for energy slackens, ATP produced by catabolism is used to replenish the PCr reservoir by reversal of the creatine kinase reaction. Organisms in the lower phyla employ other PCr-like molecules (collectively called phosphagens) as phosphoryl reservoirs. 

Kinases Direct transfer of terminal phosphoryl group of ATP to substrate Creatine kinase Adenylate kinase Hexokinasc Phosphoglycerate kinase Pyruvate kinase Protein kinase Myokinase Phosphofructokinase Type 1 (M-S-E) Type 1 Type 1 Type 1 Type 2 (S-M-E)... 

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