Thymidine monophosphate biosynthesis

In Fig. 1 various targets of some important cytostatic agents are depicted. Their main mechanisms of action can be briefly summarized as follows. Pentostatin blocks purine nucleotides by inhibiting adenosine deaminase. 6-Mercaptopurine and 6-thioguanine inhibit purine ring biosynthesis and they inhibit nucleotide interconversions. Methotrexate by inhibiting dihydrofolate reduction blocks thymidine monophosphate and purine synthesis. 5-Fluorouracil also blocks thymidine monophosphate synthesis. Dactinomycin, daunorubicin, doxorubicin and mitoxantrone intercalate with DNA and inhibit RNA synthesis. L-asparaginase deaminates... 

Thymidylate synthase (TS) is the enzyme that converts 2-deoxyuridine monophosphate into thymidine monophosphate. This is a key step in the biosynthesis of DNA. This enzymatic reaction of methylation involves the formation of a ternary complex between the substrate, the enzyme, and tetrahydrofolic acid (CH2FAH4). The catalytic cycle involves the dissociation of this complex and the elimination of FAH4. It is initiated by pulling out the proton H-5, thus generating an exocyclic methylene compound. As the release of a F" " ion is energetically forbidden, the fluorine atom that replaces the proton H-5 cannot be pulled out by the base. This leads to inhibition of the enzyme (Figure 7.2). 

One-carbon moieties of different redox states are utilized in the biosynthesis of the purine nucleotides and thymidine monophosphate (also known as thymidylate), the... 

FIGURE 1. Biochemical pathways leading to the synthesis and utilization of thymidine monophosphate (TMP). TMP biosynthesis is effected both by the folate-dependent enzyme thymidylate synthetase (TS) and by thymidine kinase (TK). Using inhibitors of folate metabolism or lethal analogues of thymidine, the TS and TK pathways can be controlled to select for TK-competent or TK-deficient cells. For details, see text. 

The sugar nucleotides (an uninformative name that has been used for glycosyl nucleotides, or more strictly, glycosyl esters of nucleoside di- or mono-phosphates) were discussed in this Series12 in 1973. Since then, accumulation of new data about these derivatives has continued, and now, about 35 representatives of this class are known to participate in the biosynthesis of polysaccharide chains of bacterial polymers (for a survey, see Ref. 13). These include glycosyl esters of uridine 5 -diphosphate (UDP), thymidine 5 -diphosphate (dTDP), guanosine 5 -diphosphate (GDP), cytidine 5 -diphosphate (CDP), cytidine 5 -monophosphate (CMP), and adenosine 5 -diphosphate (ADP). 

Kornberg s work on the biosynthesis of deoxyribonucleic acid has shown that enzymes in Escherichia coli extracts catalyze the formation of the 5-triphosphates of 2-deoxyadenosine, 2-deoxyguanosine, 2-deoxycyti-dine, and thymidine from the corresponding monophosphates in the presence of adenosine 5-triphosphate, but fail to catalyze phosphorylation of deoxyuridine 5-phosphate this finding could explain why uracil is not a constituent of deoxyribonucleic acid. 

Uridine diphosphate galactose (109) was prepared using seven enzymes involved in three biosynthetic pathways, immobilised on super-bead columns. This method, which converted 50% of uracyl monophosphate into UDP-galactose (109), was superior to the solution approach as enzyme stability was improved. To study the biosynthesis of the pseudosaccaharide acarbose, thymidine 5 -diphospho-4-amino-4,6-dideoxy-a-D-glucopyranose(110) was synthesised from galactose in sixteen steps. 

The Ts used for the biosynthesis of DNA are synthesized from Us by thymidylate synthase, an enzyme that requires N, lV -methylene-THF as a coenzyme. The actual substrate is dUMP (2 -deoxyuridine 5 -monophosphate) and the product is dTMP (2 -deoxy thymidine 5 -monophosphate). 

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