Identification of aspartic acid-203 in human thymidine phosphorylase as an important residue for both catalysis and non-competitive inhibition by the small molecule “crystallization chaperone” 5′-O-tritylinosine (KIN59)

Bronckaers, Annelies; Aguado, Leire; Negri, Ana; Camarasa, Marı́a-José; Balzarini, Jan; Pérez-Pérez, M J; Gago, Federico; Liekens, Sandra

doi:10.1016/j.bcp.2009.04.011

Cited by 16 publications

(18 citation statements)

References 49 publications

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“…TYMP metabolites including thymidine, 2DDR and 2DDRP did not affect agonist-induced platelet aggregation (SFIV), suggesting that the phosphate and the nucleoside-binding sites of TYMP are not responsible for its effect on platelet function. We thus examined whether another element of the active site of human TYMP, as partic acid-203 27 , affected its association with SFKs using a potent Asp203-binding TYMP inhibitor, KIN59 28,29 . As shown in Figure 4A, collagen stimulation significantly increased the amount of TYMP that co-precipitated with Lyn.…”

Section: Resultsmentioning

confidence: 99%

Thymidine Phosphorylase Participates in Platelet Signaling and Promotes Thrombosis

Gigante

Pérez‐Pérez

et al. 2014

Circulation Research

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Rationale Platelets contain abundant thymidine phosphorylase (TYMP), which is highly expressed in diseases with high risk of thrombosis, such as atherosclerosis and type II diabetes. Objective Test the hypothesis that TYMP participates in platelet signaling and promotes thrombosis. Methods and Results By using a ferric chloride (FeCl3) induced carotid artery injury thrombosis model, we found time to blood flow cessation was significantly prolonged in Tymp−/− and Tymp+/− mice compared to wild type (WT) mice. Bone marrow transplantation and platelet transfusion studies demonstrated that platelet TYMP was responsible for the antithrombotic phenomenon in the TYMP deficient mice. Collagen-, collagen-related peptide (CRP)-, adenosine diphosphate-and/or thrombin-induced platelet aggregation were significantly attenuated in Tymp+/− and Tymp−/− platelets, and in WT or human platelets pretreated with TYMP inhibitor KIN59. Tymp deficiency also significantly decreased agonist-induced P-select in expression. TYMP contains an N-terminal SH3 domain binding proline-rich motif and forms a complex with the tyrosine kinases Lyn, Fyn and Yes in platelets. TYMP-associated Lyn was inactive in resting platelets, and TYMP trapped and diminished active Lyn after collagen stimulation. Tymp/Lyn double haploinsufficiency diminished the antithrombotic phenotype of Tymp+/− mice. TYMP deletion or inhibition of TYMP with KIN59 dramatically increased PECAM-1 tyrosine phosphorylation and diminished CRP or collagen induced AKT phosphorylation. In vivo administration of KIN59 significantly inhibited FeCl3 induced carotid artery thrombosis without affecting hemostasis. Conclusion TYMP participates in multiple platelet signaling pathways and regulates platelet activation and thrombosis. Targeting TYMP might be a novel anti-platelet and anti-thrombosis therapy.

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Section: Resultsmentioning

confidence: 99%

Thymidine Phosphorylase Participates in Platelet Signaling and Promotes Thrombosis

Gigante

Pérez‐Pérez

et al. 2014

Circulation Research

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“…Limited studies related TP to angiogenesis (De Bruin et al , 2006; Bronckaers et al , 2009a, 2009b). From these studies, it was concluded that TP expression and activity was clearly related to the presence of angiogenesis.…”

Section: Discussionmentioning

confidence: 99%

“…To study the role of TP in angiogenesis, L-dR is often used (Nakajima et al , 2009), including other inhibitors of TP (Liekens et al , 2007; Bronckaers et al , 2009a, 2009b). Enzymatic inhibition of TP, inhibited the induction of angiogenesis, indicating that the enzymatic activity of TP in indispensable for these effects.…”

Section: Discussionmentioning

confidence: 99%

“…Numerous immunohistochemical and TP-activity studies have shown an increased TP expression and activity in a wide-range of solid tumours, compared with normal healthy tissues (Ackland and Peters, 1999; De Bruin et al , 2006; Bronckaers et al , 2009a, 2009b). A high TP expression in these tumour sites has clearly been related to a high-microvessel density, the induction of metastasis and a poor prognosis for the patient (Kidd et al , 2005; Nakayama et al , 2005; Hayashi et al , 2008).…”

mentioning

confidence: 99%

“…From these studies about TP, we conclude that TP is involved in two different, but overlapping actions. First, it is related to a higher invasive property and metastatic potential of the cancer cells (De Bruin et al , 2006; Yu et al , 2008; Bronckaers et al , 2009a, 2009b). Second, it is related to an increased angiogenesis.…”

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confidence: 99%

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Thymidine phosphorylase in cancer cells stimulates human endothelial cell migration and invasion by the secretion of angiogenic factors

et al. 2011

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Background:Thymidine phosphorylase (TP) is often overexpressed in tumours and has a role in tumour aggressiveness and angiogenesis. Here, we determined whether TP increased tumour invasion and whether TP-expressing cancer cells stimulated angiogenesis.Methods:Angiogenesis was studied by exposing endothelial cells (HUVECs) to conditioned medium (CM) derived from cancer cells with high (Colo320TP1=CT-CM, RT112/TP=RT-CM) and no TP expression after which migration (wound-healing-assay) and invasion (transwell-assay) were determined. The involvement of several angiogenic factors were examined by RT–PCR, ELISA and blocking antibodies.Results:Tumour invasion was not dependent on intrinsic TP expression. The CT-CM and RT-CM stimulated HUVEC-migration and invasion by about 15 and 40%, respectively. Inhibition by 10 μ TPI and 100 μ L-dR, blocked migration and reduced the invasion by 50–70%. Thymidine phosphorylase activity in HUVECs was increased by CT-CM. Reverse transcription-polymerase chain reaction revealed a higher mRNA expression of bFGF (Colo320TP1), IL-8 (RT112/TP) and TNF-α, but not VEGF. Blocking antibodies targeting these factors decreased the migration and invasion that was induced by the CT-CM and RT-CM, except for IL-8 in CT-CM and bFGF in RT-CM.Conclusion:In our cell line panels, TP did not increase the tumour invasion, but stimulated the migration and invasion of HUVECs by two different mechanisms. Hence, TP targeting seems to provide a potential additional strategy in the field of anti-angiogenic therapy.

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The next ten stories on antiviral drug discovery (part E): advents, advances, and adventures

Clercq¹

2010

Medicinal Research Reviews

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This review article presents the fifth part (part E) in the series of stories on antiviral drug discovery. The ten stories belonging to this fifth part are dealing with (i) aurintricarboxylic acid; (ii) alkenyldiarylmethanes; (iii) human immunodeficiency virus (HIV) integrase inhibitors; (iv) lens epithelium-derived growth factor as a potential target for HIV proviral DNA integration; (v) the status presens of neuraminidase inhibitors NAIs in the control of influenza virus infections; (vi) the status presens on respiratory syncytial virus inhibitors; (vii) tricyclic (1,N-2-ethenoguanine)-based acyclovir and ganciclovir derivatives; (viii) glycopeptide antibiotics as antivirals targeted at viral entry; (ix) the potential (off-label) use of cidofovir in the treatment of polyoma (JC and BK) virus infections; and (x) finally, thymidine phosphorylase as a target for both antiviral and anticancer agents.

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Identification of aspartic acid-203 in human thymidine phosphorylase as an important residue for both catalysis and non-competitive inhibition by the small molecule “crystallization chaperone” 5′-O-tritylinosine (KIN59)

Cited by 16 publications

References 49 publications

Thymidine Phosphorylase Participates in Platelet Signaling and Promotes Thrombosis

Thymidine Phosphorylase Participates in Platelet Signaling and Promotes Thrombosis

Thymidine phosphorylase in cancer cells stimulates human endothelial cell migration and invasion by the secretion of angiogenic factors

The next ten stories on antiviral drug discovery (part E): advents, advances, and adventures

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