Enzymes reductase kinase

This enzyme [EC 2.7.1.109], also known simply as reductase kinase, catalyzes the reaction of ATP with the enzyme 3-hydroxy-3-methylglutaryl-CoA reductase (NADPH) producing ADP and the phosphorylated form of the reductase. This phosphorylation inactivates the reductase. Histones can substitute for the reductase as substrates. 

PYRUVATE CARBOXYLASE PYRUVATE DEHYDROGENASE KINASE PYRUVATE KINASE PYRUVATE, WATER DIKINASE RESTRICTION ENZYMES RHODOPSIN KINASE RIBOFLAVIN KINASE RIBONUCLEOTIDE REDUCTASE... 

Gemcitabine is phosphorylated initially by the enzyme deoxycytidine kinase and then by other nucleoside kinases to the di- and triphosphate nucleotide forms, which then inhibit DNA synthesis. Inhibition is considered to result from two actions inhibition of ribonucleotide reductase by gemcitabine diphosphate, which reduces the level of deoxyribonucleoside triphosphates required for the synthesis of DNA and incorporation of gemcitabine triphosphate into DNA. Following incorporation of gemcitabine nucleotide, only one additional nucleotide can be added to the growing DNA strand, resulting in chain termination. 

Glucagon decreases cholesterol synthesis in isolated hepatocytes [131,132] apparently because it reduces the fraction of hydroxymethylglutaryl-CoA reductase in the active form [131,132], This is due to an increase in reductase kinase activity [133], However, there is no evidence that cAMP-dependent protein kinase phos-phorylates either the reductase, reductase kinase or reductase kinase kinase [134], It has been proposed that the phosphorylation state of these enzymes is indirectly controlled through changes in the activity of protein phosphatase I [132,134], This phosphatase can dephosphorylate and activate the reductase [134,135] and its activity can be controlled by a heat stable inhibitor (inhibitor 1), the activity of which is increased by cAMP-dependent phosphorylation [136,137], Since the phosphorylated forms of acetyl-CoA carboxylase, ATP-citrate lyase, pyruvate kinase, phos-phorylase, phosphorylase kinase and glycogen synthase are also substrates for protein phosphatase I [135], this mechanism could also contribute to their phosphorylation by glucagon. 

Figure 6.5 Regulation of HMG-CoA reductase. HMG-CoA reductase is active in the dephospho-rylated state phosphorylation (inhibition) is catalysed by reductase kinase, an enzyme whose activity is also regulated by phosphorylation by reductase kinase kinase. Hormones such as glucagon and adrenalin (epinephrine) negatively affect cholesterol biosynthesis by increasing the activity of the inhibitor of phosphoprotein phosphatase-1, PPI-1, (by raising cAMP levels) and so reducing the activation of HMG-CoA reductase. Conversely, insulin stimulates the removal of phosphates (and lowers cAMP levels), and thereby activates HMG-CoA reductase activity. Additional regulation of HMG-CoA reductase occurs through an inhibition of synthesis of the enzyme by elevation in intracellular cholesterol levels.

AMP-dependent protein kinase is an enzyme that binds AMP and can phosphorylate both acetyl-CoA carboxylase and HMG-CoA reductase kinase. In each case, the enzyme is inactivated. 

An ADA-resistant purine analog, cladribine (2-chlorodeoxyadenosine 2-CdA) has demonstrated potent activity in hairy cell leukemia, CLL, and low-grade lymphomas. After intracellular phosphorylation by deoxycytidine kinase and conversion to cladribine triphosphate, it is incorporated into DNA. It produces DNA strand breaks and depletion of NAD and adenosine triphosphate (ATP), as well as apoptosis, and is a potent inhibitor of ribonucleotide reductase. The drug does not require cell division to be cytotoxic. Resistance is associated with loss of the activating enzyme, deoxycytidine kinase, or escape of ribonucleotide reductase from inhibition. 

Beg, Z.H. Stonik, J.A. Reversible inactivation of 3-hydroxy-3-metbylglutar-yl coenzyme A reductase reductase kinase and mevalonate kinase are separate enzymes. Biochem. Biophys. Res. Commun., 108, 559-566 (1982)... 

Beg, Z.H. Stonik, J.A. Brewer, B. Characterization and regulation of reductase kinase, a protein kinase that modulates the enzymic activity of 3-hy-droxy-3-methylglutaryl-coenzyme A reductase. Proc. Natl. Acad. Sci. USA, 76, 4375-4379 (1979)... 

MVA is phosphorylated in two steps to the mono- and the diphosphate (MVAP and MVAPP), by the specific ATP-dependent enzymes MVA kinase (ATP-mevalonate-phosphotransferase, EC 2.7.1.36) and MVAP kinase (EC 2.7.4.2), respectively. MVAPP is converted into IPP by a decarboxylase (EC 4.1.1.33). The kinases and the decarboxylase have not yet been given as much attention as HMG-CoA reductase, and have only been characterized in a few plant species. Recently, MVA kinase was partially purified from C. roseus suspension cultures. The enzyme proved to be unstable and was present only at low activity levels (90). 

In HeLa ceils hydroxyurea is an efficient inhibitor of histone synthesis. This action requires protein synthesis and leads to rapid disappearance of cytoplasmatic histone mRNA The effect is not specific for hydroxyurea since suppression of DNA synthesis by arabino-cytosine or temperature-sensitive mutations yields analogous results. Similarly, the synthesis of some enzymes necessary for DNA replication and active in S-phase is altered by hydroxyurea. Increased activity of ribonucleotide reductase in HeLa and in hamster cells and of the salvage enzyme thymidine kinase in HeLa cells and KB cells has been observed, probably as a consequence of the increased fraction of cells in S-phase. Repression occurs for thymidine kinase in human lymphocytes and for ornithine decarboxylase in Chinese hamster fibroblasts whereas no or only slight effects were seen on ribonucleotide reductase in hamster fibroblasts , on thymidylate synthase in extracts from synchronous mouse cells " , and on DNA polymerase in rabbit kidney cells or HeLa cells . ... 

HMG-CoA reductase is also regulated by a reversible phosphorylation/ dephosphorylation cycle (Figure 7.17). The phosphorylated reductase is inactive and the amounts of the phosphorylated enzyme can be shown to be increased when its activity is decreased by mevalonate or glucagon. The protein kinase which inactivates HMG-CoA reductase is itself subject to phosphorylation. Interestingly, both ATP and ADP are needed. Apparently ADP binds to a different site on the reductase kinase and acts as an allosteric effector. The phosphatases which activate HMG-CoA reductase are highly sensitive to NaF. Both the kinase and phosphatases are present in both microsomal and cytoplasmic fractions. 

Deoxynucleotides for DNA synthesis are made at the nucleoside diphosphate level and then have to be phosphorylated up to the triphosphate using a kinase and ATP. The reducing equivalents for the reaction come from a small protein, thioredoxin, that contains an active site with two cysteine residues. Upon reduction of the ribose to the 2 -deoxyri-bose, the thioredoxin is oxidized to the disulfide. The thioredoxin(SS) made during the reaction is recycled by reduction with NADPH by the enzyme thioredoxin reductase. 

Ribonucleotide reductase works on ribo-A, -U, -G, -C diphosphates to give the deoxynucleotide. The deoxyuridine, which is useless for RNA synthesis, is converted to deoxythymidine by the enzyme thymidylate synthase, which uses methylene tetrahydrofolate as a one-carbon donor. The odd thing here is that ribonucleotide reductase uses the UDP as a substrate to give the dUDP. This must then be hydrolyzed to the dUMP before thymidylate synthase will use it to make dTMP. Then the dTMP has to be kinased (phosphorylated) up to dTTP before DNA can be made. 

The phosphoribosyltiansferases The nucleoside kinases Nucleoside phosphokinases Nucleotide reductases Methylases and demethylases Other anabolic enzymes Catabolism... 

ADP as a substrate in enzyme reactions, ADENYLATE KINASE (or MYOKINASE) ATP SYNTHASE CREATINE KINASE NUCLEOSIDE DIPHOSPHATE KINASE PHOSPHOGLYCERATE KINASE PYRUVATE KINASE RIBONUCLEOTIDE REDUCTASE SULFATE ADENYLYLTRANSFERASE (ADP) [ADP]/[ATP] ratio,... 

Suramin (Germanin) is a derivative of a nonmetallic dye whose antiparasitic mechanism of action is not clear. It appears to act on parasite specific a-glyc-erophosphate oxidase, thymidylate synthetase, dihydrofolate reductase, and protein kinase but not on host enzymes. 

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