Abstract：
:Catabolism of coagulation factor VIII (FVIII) is mediated by low-density lipoprotein receptor-related protein (LRP). The ligand-binding sites of LRP are formed by complement-type repeats (CR), and CR clusters II and IV bind most of LRP ligands. FVIII contains two major LRP-binding sites located in the A2 and A3 domains. This study was aimed to identify specific complement-type repeats of LRP involved in interaction with the A2 site and to probe their functional importance in A2 catabolism. We generated individual LRP clusters II, III and IV, along with nine overlapping CR triplets encompassing clusters II and IV in a baculovirus expression system and studied their interaction with isolated A2. In surface plasmon resonance (SPR) assay, A2 bound to clusters II and IV with KDs 22 and 39 nM, respectively, and to the majority of CR triplets with affinities in the range of KDs 25-90 nM. Similar affinities were determined for A2 interaction with a panel of CR doublets overlapping cluster II (CR 3-4, 4-5, 5-6, 6-7 and 7-8). These LRP fragments inhibited the binding of 125I-A2 to LRP in solid-phase assay, LRP-mediated internalization of 125I-A2 in cell culture and 125I-A2 clearance from the mouse circulation. Point mutations of critical A2 residues of the LRPbinding site resulted in differential reduction or abolishment of its binding to LRP fragments. We conclude that A2 interacts with LRP via multiple binding sites spanning CR 3-8 in cluster II and CR 23-29 in cluster IV, and the minimal A2-binding unit of LRP is formed by two adjacent CR.
journal_name
Thromb Haemostjournal_title
Thrombosis and haemostasisauthors
Sarafanov AG,Makogonenko EM,Andersen OM,Mikhailenko IA,Ananyeva NM,Khrenov AV,Shima M,Strickland DK,Saenko ELsubject
Has Abstractpub_date
2007-12-01 00:00:00pages
1170-81issue
6eissn
0340-6245issn
2567-689Xpii
07121170journal_volume
98pub_type
杂志文章abstract：:The pathogenesis of abdominal aortic aneurysm (AAA) involves a central component of chronic inflammation which is predominantly mediated by myeloid cells. We hypothesized that the local inflammatory activity may be reflected in systemic alterations of neutrophil and monocyte populations as well as in soluble factors o...
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journal_title：Thrombosis and haemostasis
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journal_title：Thrombosis and haemostasis
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journal_title：Thrombosis and haemostasis
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journal_title：Thrombosis and haemostasis
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journal_title：Thrombosis and haemostasis
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journal_title：Thrombosis and haemostasis
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journal_title：Thrombosis and haemostasis
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journal_title：Thrombosis and haemostasis
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