Crystallization and X-ray Diffraction Study of Recombinant Platelet-derived Endothelial Cell Growth Factor

Spraggon, Glen; Stuart, David I.; Ponting, Chris P.; Finnis, C; Sleep, Darrell; Jones, E Y

doi:10.1006/jmbi.1993.1632

Cited by 6 publications

(3 citation statements)

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“…25 Thus, Spraggon et al reported crystals of human TP for which, despite using a synchroton X-ray source, diffraction was limited to 3.5 Å resolution. 26 Finally, in 2004, Norman et al successfully solved at 2.1 Å resolution the structure of human TP in complex with the small and potent inhibitor 5-chloro-6-[1-(2-iminopyrrolidinyl)methyl] uracil (TPI) (see Section 6.A). 25 Nonetheless, limited proteolysis with trypsin was found to be necessary and this treatment yielded a structure in which amino acids 409 and 410 were missing and the loop formed by amino acid residues 405-416 was disordered.…”

Section: Structure Of Tpmentioning

confidence: 99%

The dual role of thymidine phosphorylase in cancer development and chemotherapy

Bronckaers¹,

Gago²,

Balzarini³

et al. 2009

Medicinal Research Reviews

179

176

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Thymidine phosphorylase (TP), also known as "platelet-derived endothelial cell growth factor" (PD-ECGF), is an enzyme, which is upregulated in a wide variety of solid tumors including breast and colorectal cancers. TP promotes tumor growth and metastasis by preventing apoptosis and inducing angiogenesis. Elevated levels of TP are associated with tumor aggressiveness and poor prognosis. Therefore, TP inhibitors are synthesized in an attempt to prevent tumor angiogenesis and metastasis. TP is also indispensable for the activation of the extensively used 5-fluorouracil prodrug capecitabine, which is clinically used for the treatment of colon and breast cancer. Clinical trials that combine capecitabine with TP-inducing therapies (such as taxanes or radiotherapy) suggest that increasing TP expression is an adequate strategy to enhance the antitumoral efficacy of capecitabine. Thus, TP plays a dual role in cancer development and therapy: on the one hand, TP inhibitors can abrogate the tumorigenic and metastatic properties of TP; on the other, TP activity is necessary for the activation of several chemotherapeutic drugs. This duality illustrates the complexity of the role of TP in tumor progression and in the clinical response to fluoropyrimidine-based chemotherapy.

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Section: Structure Of Tpmentioning

confidence: 99%

The dual role of thymidine phosphorylase in cancer development and chemotherapy

Bronckaers¹,

Gago²,

Balzarini³

et al. 2009

Medicinal Research Reviews

179

176

View full text Add to dashboard Cite

show abstract

“…Indeed, well diffracting crystals of HTP have been reported to be difficult to grow. Thus Spraggon et al [30] have reported HTP crystals for which diffraction was limited to 3.5 Å resolution. Norman et al [27] were successful in solving the HTP structure to 2.1 Å resolution, although the enzyme required partial digestion, resulting in several disordered residues.…”

Section: Ligand Bindingmentioning

confidence: 99%

Structural basis for non-competitive product inhibition in human thymidine phosphorylase: implications for drug design

Omari

Bronckaers

Liekens

et al. 2006

Biochemical Journal

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HTP (human thymidine phosphorylase), also known as PD-ECGF (platelet-derived endothelial cell growth factor) or gliostatin, has an important role in nucleoside metabolism. HTP is implicated in angiogenesis and apoptosis and therefore is a prime target for drug design, including antitumour therapies. An HTP structure in a closed conformation complexed with an inhibitor has previously been solved. Earlier kinetic studies revealed an ordered release of thymine followed by ribose phosphate and product inhibition by both ligands. We have determined the structure of HTP from crystals grown in the presence of thymidine, which, surprisingly, resulted in bound thymine with HTP in a closed dead-end complex. Thus thymine appears to be able to reassociate with HTP after its initial ordered release before ribose phosphate and induces the closed conformation, hence explaining the mechanism of non-competitive product inhibition. In the active site in one of the four HTP molecules within the crystal asymmetric unit, additional electron density is present. This density has not been previously seen in any pyrimidine nucleoside phosphorylase and it defines a subsite that may be exploitable in drug design. Finally, because our crystals did not require proteolysed HTP to grow, the structure reveals a loop (residues 406-415), disordered in the previous HTP structure. This loop extends across the active-site cleft and appears to stabilize the dimer interface and the closed conformation by hydrogen-bonding. The present study will assist in the design of HTP inhibitors that could lead to drugs for anti-angiogenesis as well as for the potentiation of other nucleoside drugs.

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“…As the name implies, it was thought to be a classic polypeptide growth factor that would bind to a cognate cell-surface receptor and elicit a cellular response. PD-ECGF was first purified to homogeneity in 1989 [15] and its crystal structure elucidated, though not to a particularly high resolution [16]. The PD-ECGF gene was isolated [17].…”

Section: Platelet-derived Endothelial-cell Growth Factormentioning

confidence: 99%

Thymidine phosphorylase, 2-deoxy-D-ribose and angiogenesis

Brown

Bicknell

1998

Biochemical Journal

196

157

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Angiogenesis is the term used to describe the formation of new blood vessels from the existing vasculature. In order to attract new vessels, a tissue must release an endothelial-cell chemoattractant. 2-Deoxy-D-ribose is produced in vivo by the catalytic action of thymidine phosphorylase (TP) on thymidine and has recently been identified as an endothelial-cell chemoattractant and angiogenesis-inducing factor. TP, previously known only for its role in nucleotide salvage, is now known to be angiogenic. TP expression is elevated in many solid tumours and in chronically inflamed tissues, both known areas of active angiogenesis. There is evidence that TP is also involved in physiological angiogenesis such as endometrial angiogenesis during the menstrual cycle. The majority of known endothelial-cell chemoattractants are polypeptides that bind to endothelial-cell-surface receptors. In contrast, 2-deoxy-D-ribose appears to lack a cell-surface receptor. Glucose is another sugar that acts as an endothelial-cell chemoattractant. The migratory activity of glucose is blocked by ouabain. It is possible that 2-deoxy-D-ribose and glucose stimulate endothelial-cell migration via a similar mechanistic pathway.

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Crystallization and X-ray Diffraction Study of Recombinant Platelet-derived Endothelial Cell Growth Factor

Cited by 6 publications

References 0 publications

The dual role of thymidine phosphorylase in cancer development and chemotherapy

The dual role of thymidine phosphorylase in cancer development and chemotherapy

Structural basis for non-competitive product inhibition in human thymidine phosphorylase: implications for drug design

Thymidine phosphorylase, 2-deoxy-D-ribose and angiogenesis

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