Ancylostoma caninum Anticoagulant Peptide Blocks Metastasis In Vivo and Inhibits Factor Xa Binding to Melanoma Cells In Vitro
Ancylostoma caninum Anticoagulant Peptide Blocks Metastasis In Vivo and Inhibits Factor Xa Binding to Melanoma Cells In Vitro
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犬钩虫抗凝肽可阻断体内转移并在体外抑制 Xa 因子与黑色素瘤细胞的结合
DOI：
10.1055/s-0037-1615117
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发表时间：
1998
影响因子：
6.7
通讯作者：
M. Cappello
中科院分区：
文献类型：
--
作者：
K. Donnelly;M. Bromberg;A. Milstone;J. McNiff;G. Terwilliger;William K. Konigsberg;M. Cappello
我们评价了重组犬钩虫抗凝肽（rAcAP）的体内抗转移活性，rAcAP是一种有效的（Ki = 265 pM）和特异性活性位点的人凝血因子Xa抑制剂，最初分离自吸血钩虫。在尾静脉注射LOX人黑素瘤细胞之前，用rAcAP（0.01- 0.2mg/小鼠）皮下注射SCID小鼠导致肺转移的剂量依赖性减少。为了阐明rAcAP的抗转移活性的潜在机制，进行实验以鉴定因子Xa和LOX之间的特异性相互作用。生物素化因子Xa与LOX单层的结合是特异性的和可饱和的（Kd = 15 nM）。使用先前鉴定的因子Xa结合蛋白（包括因子V/Va、效应细胞蛋白酶受体-1和组织因子途径抑制剂）的抗体的竞争实验未能暗示这些分子中的任何分子为因子Xa的显著结合位点。LOX也支持功能性凝血酶原酶活性，在Xa因子浓度为2.4 nM时检测到凝血酶生成的一半最大速率。使用过量的rAcAP或活性位点阻断因子Xa（EGR-Xa）的另外的竞争实验揭示，大部分总因子Xa与LOX的结合是通过与酶的活性位点的相互作用介导的，预测绝大多数细胞相关因子Xa不直接参与凝血酶生成。除了建立两种不同的机制因子Xa结合黑色素瘤，这些数据提出的可能性，rAcAP的抗转移作用在体内可能涉及新的非凝血途径，也许通过抑制活性位点介导的因子Xa和肿瘤细胞之间的相互作用。
Summary We evaluated the in vivo anti-metastatic activity of recombinant Ancylostoma caninum Anticoagulant Peptide (rAcAP), a potent (Ki = 265 pM) and specific active site inhibitor of human coagulation factor Xa originally isolated from bloodfeeding hookworms. Subcutaneous injection of SCID mice with rAcAP (0.01-0.2 mg/mouse) prior to tail vein injection of LOX human melanoma cells resulted in a dose dependent reduction in pulmonary metastases. In order to elucidate potential mechanisms of rAcAP’s anti-metastatic activity, experiments were carried out to identify specific interactions between factor Xa and LOX. Binding of biotinylated factor Xa to LOX monolayers was both specific and saturable (Kd = 15 nM). Competition experiments using antibodies to previously identified factor Xa binding proteins, including factor V/Va, effector cell protease receptor-1, and tissue factor pathway inhibitor failed to implicate any of these molecules as significant binding sites for Factor Xa. Functional prothrombinase activity was also supported by LOX, with a half maximal rate of thrombin generation detected at a factor Xa concentration of 2.4 nM. Additional competition experiments using an excess of either rAcAP or active site blocked factor Xa (EGR-Xa) revealed that most of the total factor Xa binding to LOX is mediated via interaction with the enzyme’s active site, predicting that the vast majority of cell-associated factor Xa does not participate directly in thrombin generation. In addition to establishing two distinct mechanisms of factor Xa binding to melanoma, these data raise the possibility that rAcAP’s antimetastatic effect in vivo might involve novel non-coagulant pathways, perhaps via inhibition of active-site mediated interactions between factor Xa and tumor cells.
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效应细胞蛋白酶受体1的鉴定。
DOI：
--
发表时间：
1990
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Altieri,DC;Edgington,TS
通讯作者：
Edgington,TS
DOI：
10.1016/s0021-9258(19)49691-9
发表时间：
1992-07
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
S. Rabbani;A. Mazar;S. Bernier;M. Haq;I. Bolivar;J. Henkin;David Goltzman
通讯作者：
S. Rabbani;A. Mazar;S. Bernier;M. Haq;I. Bolivar;J. Henkin;David Goltzman
影响因子：
1.5
作者：
Cappello, M;Hawdon, JM;Hotez, PJ
通讯作者：
Hotez, PJ
DOI：
10.1073/pnas.92.18.8205
发表时间：
1995-08-29
影响因子：
11.1
作者：
BROMBERG, ME;KONIGSBERG, WH;GAREN, A
通讯作者：
GAREN, A
高分子量激肽原仅与内皮细胞膜结合，影响血管内皮的活化。
影响因子：
20.3
作者：
Hasan,AA;Cines,DB;Ngaiza,JR;Jaffe,EA;Schmaier,AH
通讯作者：
Schmaier,AH