Atomic Force Microscopy-Based Molecular Recognition of a Fibrinogen Receptor on Human Erythrocytes
Atomic Force Microscopy-Based Molecular Recognition of a Fibrinogen Receptor on Human Erythrocytes
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DOI：
10.1021/nn1009648
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发表时间：
2010-08-01
期刊：
影响因子：
17.1
通讯作者：
Santos, Nuno C.
中科院分区：
文献类型：
--
作者：
Carvalho, Filomena A.;Connell, Simon;Santos, Nuno C.
红细胞聚集性增加与心血管疾病发生率增加的既定假设是基于血浆黏附蛋白浓度的增加，特别是纤维蛋白原。纤维蛋白原诱导的红细胞聚集被认为是由于其与红细胞膜的非特异性结合而引起的。相比之下，已知的是，血小板表面表达一种纤维蛋白原整合素受体(膜糖蛋白复合体α(IIb)β(3))。通过原子力显微镜(AFM)的力光谱测量，我们证明了纤维蛋白原与红细胞膜上的未知受体之间存在单分子相互作用，与血小板结合相比亲和力较低(平均纤维蛋白原红细胞结合力和血小板平均结合力分别为79和97pN)。这种受体不像血小板受体那样受到钙和依替非那肽(一种AMP特异性抑制剂)的强烈影响。然而，它被表菲拉肽抑制，表明它是一种与α(II)β(3)相关的整合素。对Glanzmann血栓症(一种罕见的遗传性出血性疾病，由α(II)β(3)缺乏引起)患者的结果显示(第一次)纤维蛋白原红细胞结合受损。与基因测序数据的相关性表明，红细胞上纤维蛋白原受体的一个单位是β(3)基因表达的产物，该基因在该患者中被发现突变。这项工作证明并验证了基于AFM的力谱作为一种高度敏感、快速和低操作成本的纳米工具的适用性，用于诊断导致血液病的基因突变，并对其严重性进行无偏见的功能评估。
The established hypothesis for the increase on erythrocyte aggregation associated with a higher incidence of cardiovascular pathologies is based on an increase on plasma adhesion proteins concentration, particularly fibrinogen. Fibrinogen-induced erythrocyte aggregation has been considered to be caused by its nonspecific binding to erythrocyte membranes. In contrast, platelets are known to have a fibrinogen integrin receptor expressed on the membrane surface (the membrane glycoprotein complex alpha(IIb)beta(3)). We demonstrate, by force spectroscopy measurements using an atomic force microscope (AFM), the existence of a single molecule interaction between fibrinogen and an unknown receptor on the erythrocyte membrane, with a lower but comparable affinity relative to platelet binding (average fibrinogen erythrocyte and platelet average (un)binding forces were 79 and 97 pN, respectively). This receptor is not as strongly influenced by calcium and eptifibatide (an amp) specific inhibitor) as the platelet receptor. However, its inhibition by eptifibatide indicates that it is an alpha(II)beta(3)-related integrin. Results obtained for a Glanzmann thrombastenia (a rare hereditary bleeding disease caused by alpha(II)beta(3) deficiency) patient show (for the first time) an impaired fibrinogen erythrocyte binding. Correlation with genetic sequencing data demonstrates that one of the units of the fibrinogen receptor on erythrocytes is a product of the expression of the beta(3) gene, found to be mutated in this patient. This work demonstrates and validates the applicability of AFM-based force spectroscopy as a highly sensitive, rapid and low operation cost nanotool for the diagnostic of genetic mutations resulting in hematological diseases, with an unbiased functional evaluation of their severity.