Thymidine monophosphate kinase (TMPK) of Mycobacterium tuberculosis (TMPKmt) represents an attractive target for blocking the bacterial DNA synthesis. In an attempt to find high-affinity inhibitors of TMPKmt, a cavity in the enzyme at the 3'-position was explored via the introduction of various substituents at the 3'-position of the thymidine monophosphate (dTMP) scaffold. Various 3'-C-branched...

The enzymes deoxycytidylate deaminase (EC) and thymidylate synthase (EC) are functionally associated with one another, since they catalyze sequential reactions. In T4 coliphage infection the two enzymes are found in dNTP synthetase, a multienzyme complex for deoxyribonucleotide biosynthesis. Protein-protein interactions involving the phage-coded forms of these two enzymes have been explored in ...

Here we report on a Chlamydia trachomatis gene that complements the growth defect of a thymidylate synthase-deficient strain of Escherichia coli. The complementing gene encodes a 60.9-kDa protein that shows low level primary sequence homology to a new class of thymidylate-synthesizing enzymes, termed flavin-dependent thymidylate synthases (FDTS). Purified recombinant chlamydial FDTS (CTThyX) co...

Gene silencing and RNA interference are major cellular processes that control gene expression via the cleavage of target mRNA. Eukaryotic translation initiation factor 2C2 (EIF2C2, Argonaute protein 2, Ago2) is considered to be the major player of RNAi as it is the core component of RISC complexes. While a considerable amount of research has focused on RNA interference and its associated mechan...

AMP: Deaminase Adenosine Monophosphate Deaminase; ATP 7B: Membrane-Bound Copper-Binding Adenosine Triphosphatase; cGMP: Guanosine 3’, 5’ Cyclic Monophosphate; DC: Dendritic Cells; DNA: Deoxyribonucleic Acid; dTMP: Deoxythymidine 5’ Mono-Phosphate; HL-60: Human promyelocytic Leukemia cell line; HUT-78: Human malignant lymphoblastoid cell line of Th0 phenotype; ICAM-1: Soluble Intercellar Adhesio...

We design here new nanomolar antituberculotics, inhibitors of Mycobacterium tuberculosis thymidine monophosphate kinase (TMPKmt), by means of structure-based molecular design. 3D models of TMPKmt-inhibitor complexes have been prepared from the crystal structure of TMPKmt cocrystallized with the natural substrate deoxythymidine monophosphate (dTMP) (1GSI) for a training set of 15 thymidine analo...

Thymidylate synthase (TS) catalyzes the transfer of a methyl group from methylenetetrahydrofolate to dUMP to form dTMP. It is a primary target in the chemotherapy of colorectal cancers and some other neoplasms. In order to obtain pure protein for analysis of structure and biological function, an expression vector TS-pET28b (+) was constructed by inserting wild-type human thymidylate synthase (h...

Folate derivatives are utilized in single-carbon transfer reactions for many critical pathways, including methionine and DNA synthesis (see Fig. 1). For example, 5,10-methylene tetrahydrofolate (5,10-methylene THF) is required for conversion of dUMP to dTMP. It can also be converted to 10formyltetrahydrofolate for the synthesis of the purine ring, or reduced to 5-methyltetrahydrofolate (5-methy...

Uracil-DNA-glycosylase (UDG) is the first enzyme in the base excision repair pathway for removal of uracil in DNA. The disturbance of reparation system results in occurrence of a number of diseases, including various kinds of a cancer. Uracil (Ura) in DNA may result from incorporation of dUMP instead of dTMP or from spontaneous deamination of cytosine in DNA. The majority of organisms delete th...

1. A mouse liver plasma-membrane preparation was solubilized in an N-dodecylsarcosinate-Tris buffer, pH7.8, and the proteins and glycoproteins were separated by a rate-zonal centrifugation in sucrose-detergent gradients. 2. A peak of alkaline phosphodiesterase activity which sedimented ahead of the 5'-nucleotidase peak was associated with a major glycoprotein component of the plasma membrane. 3...