PubMed:21840881 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PubMed/sourceid/21840881","sourcedb":"PubMed","sourceid":"21840881","source_url":"http://www.ncbi.nlm.nih.gov/pubmed/21840881","text":"Thrombomodulin domains attenuate atherosclerosis by inhibiting thrombin-induced endothelial cell activation.\nAIMS: Thrombin modulates the formation of atherosclerotic lesions by stimulating a variety of cellular effects through protease-activated receptor-1 (PAR-1) activation. Thrombomodulin (TM) inhibits thrombin effects by binding thrombin through its domains 2 and 3 (TMD23). We investigated whether recombinant TMD23 (rTMD23) could inhibit atherosclerosis via its thrombin-binding ability.\nMETHODS AND RESULTS: Wild-type mouse rTMD23 and three mutants with altered thrombin-binding sites, rTMD23 (I425A), rTMD23 (D424A/D426A), and rTMD23 (D424A/I425A/D426A), were expressed and purified in the Pichia pastoris expression system. Wild-type rTMD23 and rTMD23 (D424A/D426A) could effectively bind thrombin, activate protein C, and prolong thrombin clotting time, whereas rTMD23 (I425A) and rTMD23 (D424A/I425A/D426A) lost these functions. Wild-type rTMD23, but not rTMD23 (I425A), decreased both the thrombin-induced surface PAR-1 internalization and the increase in cytoplasmic Ca(2+) concentrations in endothelial cells (ECs). Wild-type rTMD23 and rTMD23 (D424A/D426A) also inhibited thrombin-induced adhesion molecules and monocyte chemoattractant protein-1 expression and increased permeability in ECs, whereas rTMD23 (I425A) and rTMD23 (D424A/I425A/D426A) had no such effects. Furthermore, wild-type rTMD23 and rTMD23 (D424A/D426A) were effective in reducing carotid ligation-induced neointima formation in C57BL/6 mice and atherosclerotic lesion formation in apolipoprotein E-deficient (ApoE-/-) mice, whereas rTMD23 with the I425A mutation showed impairment of this function. Wild-type rTMD23, but not rTMD23 (I425A), also markedly suppressed the PAR-1, the adhesion molecules expression, and the macrophage content in the carotid ligation model and ApoE-/- mice.\nCONCLUSION: rTMD23 protein significantly reduces atherosclerosis and neointima formation through its thrombin-binding ability.","tracks":[{"project":"Allie","denotations":[{"id":"SS1_21840881_2_0","span":{"begin":228,"end":257},"obj":"expanded"},{"id":"SS2_21840881_2_0","span":{"begin":259,"end":264},"obj":"abbr"},{"id":"SS1_21840881_3_0","span":{"begin":278,"end":292},"obj":"expanded"},{"id":"SS2_21840881_3_0","span":{"begin":294,"end":296},"obj":"abbr"},{"id":"SS1_21840881_4_0","span":{"begin":405,"end":422},"obj":"expanded"},{"id":"SS2_21840881_4_0","span":{"begin":424,"end":430},"obj":"abbr"},{"id":"SS1_21840881_8_0","span":{"begin":1107,"end":1124},"obj":"expanded"},{"id":"SS2_21840881_8_0","span":{"begin":1126,"end":1129},"obj":"abbr"}],"relations":[{"id":"AE1_21840881_2_0","pred":"abbreviatedTo","subj":"SS1_21840881_2_0","obj":"SS2_21840881_2_0"},{"id":"AE1_21840881_3_0","pred":"abbreviatedTo","subj":"SS1_21840881_3_0","obj":"SS2_21840881_3_0"},{"id":"AE1_21840881_4_0","pred":"abbreviatedTo","subj":"SS1_21840881_4_0","obj":"SS2_21840881_4_0"},{"id":"AE1_21840881_8_0","pred":"abbreviatedTo","subj":"SS1_21840881_8_0","obj":"SS2_21840881_8_0"}],"attributes":[{"subj":"SS1_21840881_2_0","pred":"source","obj":"Allie"},{"subj":"SS2_21840881_2_0","pred":"source","obj":"Allie"},{"subj":"SS1_21840881_3_0","pred":"source","obj":"Allie"},{"subj":"SS2_21840881_3_0","pred":"source","obj":"Allie"},{"subj":"SS1_21840881_4_0","pred":"source","obj":"Allie"},{"subj":"SS2_21840881_4_0","pred":"source","obj":"Allie"},{"subj":"SS1_21840881_8_0","pred":"source","obj":"Allie"},{"subj":"SS2_21840881_8_0","pred":"source","obj":"Allie"}]},{"project":"PubmedHPO","denotations":[{"id":"T1","span":{"begin":446,"end":461},"obj":"HP_0002621"}],"attributes":[{"subj":"T1","pred":"source","obj":"PubmedHPO"}]},{"project":"DisGeNET5_gene_disease","denotations":[{"id":"21840881-0#0#14#gene7056","span":{"begin":0,"end":14},"obj":"gene7056"},{"id":"21840881-0#33#48#diseaseC0003850","span":{"begin":33,"end":48},"obj":"diseaseC0003850"},{"id":"21840881-0#33#48#diseaseC0004153","span":{"begin":33,"end":48},"obj":"diseaseC0004153"}],"relations":[{"id":"0#14#gene705633#48#diseaseC0003850","pred":"associated_with","subj":"21840881-0#0#14#gene7056","obj":"21840881-0#33#48#diseaseC0003850"},{"id":"0#14#gene705633#48#diseaseC0004153","pred":"associated_with","subj":"21840881-0#0#14#gene7056","obj":"21840881-0#33#48#diseaseC0004153"}],"attributes":[{"subj":"21840881-0#0#14#gene7056","pred":"source","obj":"DisGeNET5_gene_disease"},{"subj":"21840881-0#33#48#diseaseC0003850","pred":"source","obj":"DisGeNET5_gene_disease"},{"subj":"21840881-0#33#48#diseaseC0004153","pred":"source","obj":"DisGeNET5_gene_disease"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"Allie","color":"#b5ec93","default":true},{"id":"PubmedHPO","color":"#ec93d0"},{"id":"DisGeNET5_gene_disease","color":"#93eaec"}]}]}}