Crystal structure of coagulation factor XII N-terminal domains 1-5.

IF 2.6 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS
Muhammad Saleem, Chan Li, Bubacarr G Kaira, Alexander K Brown, Monika Pathak, Shabir Najmudin, Nathan Cowieson, Ingrid Dreveny, Clare Wilson, Aleksandr Shamanaev, David Gailani, Stephanie A Smith, James H Morrissey, Helen Philippou, Jonas Emsley
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引用次数: 0

Abstract

Factor XIIa (FXIIa) is generated from its zymogen factor XII (FXII) by contact with polyanions such as inorganic polyphosphates. FXIIa cleaves the substrates prekallikrein and factor XI, triggering inflammatory cascades and plasma coagulation. From the N-terminus, FXII has fibronectin type II (FnII), epidermal growth factor-1 (EGF1), fibronectin type I (FnI), EGF2 and kringle domains. The N-terminal domains of FXII mediate polyanion and Zn2+ binding. To understand how ligand binding to polyanions and Zn2+ is coordinated across multiple domains, we determined the crystal structure of recombinant FXII domains 1-5 (FXIIHC5) to 3.4 Å resolution. A separate crystal structure of the isolated FXII FnII domain at 1.2 Å resolution revealed two bound Zn2+ ions. In FXIIHC5 a head-to-tail interaction is formed between the FnII and kringle domains, co-localizing the lysine-binding sites of the kringle domain and the cation-binding site of the FnII domain. Two FXIIHC5 monomers interlock, burying a large surface area of 2067 Å2, such that two kringle domains point outwards separated by a distance of 20 Å. The polyanion-binding site in the EGF1 domain is localized onto a plane together with the FnII and FnI domains. Using native mass spectrometry, we detected a major FXIIHC5 monomer peak and a minor dimer peak. Small-angle X-ray scattering and gel-filtration chromatography revealed the presence of monomers and dimers in solution. These FXII N-terminal domain structures provide a holistic framework to understand how the mosaic domain structure of FXII assembles diverse ligand-binding sites in three dimensions.

凝血因子XII n端结构域1-5的晶体结构。
因子XIIa (FXIIa)是由其酶原因子XII (FXII)与多阴离子(如无机多磷酸盐)接触而产生的。FXIIa切割底物prekallikin和因子XI,引发炎症级联反应和血浆凝固。从n端来看,FXII具有纤维连接蛋白II型(FnII)、表皮生长因子-1 (EGF1)、纤维连接蛋白I型(FnI)、EGF2和kringle结构域。FXII的n端结构域介导多阴离子和Zn2+的结合。为了了解配体与多阴离子和Zn2+的结合是如何跨多个结构域协调的,我们以3.4 Å分辨率确定了重组FXII结构域1-5 (FXIIHC5)的晶体结构。在1.2 Å分辨率下,分离的FXII FnII畴的单独晶体结构显示了两个结合的Zn2+离子。在FXIIHC5中,FnII和kringle结构域之间形成了头尾相互作用,kringle结构域的赖氨酸结合位点和FnII结构域的阳离子结合位点共定位。两个FXIIHC5单体互锁,掩埋了2067 Å2的大表面积,使得两个kringle域指向外,相距20 Å。EGF1结构域的多阴离子结合位点与FnII和FnI结构域一起定位在一个平面上。使用天然质谱法,我们检测到一个主要的FXIIHC5单体峰和一个次要的二聚体峰。小角x射线散射和凝胶过滤色谱显示溶液中存在单体和二聚体。这些FXII n端结构域结构为了解FXII的镶嵌结构域结构如何在三维上组装不同的配体结合位点提供了一个整体框架。
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来源期刊
Acta Crystallographica. Section D, Structural Biology
Acta Crystallographica. Section D, Structural Biology BIOCHEMICAL RESEARCH METHODSBIOCHEMISTRY &-BIOCHEMISTRY & MOLECULAR BIOLOGY
CiteScore
4.50
自引率
13.60%
发文量
216
期刊介绍: Acta Crystallographica Section D welcomes the submission of articles covering any aspect of structural biology, with a particular emphasis on the structures of biological macromolecules or the methods used to determine them. Reports on new structures of biological importance may address the smallest macromolecules to the largest complex molecular machines. These structures may have been determined using any structural biology technique including crystallography, NMR, cryoEM and/or other techniques. The key criterion is that such articles must present significant new insights into biological, chemical or medical sciences. The inclusion of complementary data that support the conclusions drawn from the structural studies (such as binding studies, mass spectrometry, enzyme assays, or analysis of mutants or other modified forms of biological macromolecule) is encouraged. Methods articles may include new approaches to any aspect of biological structure determination or structure analysis but will only be accepted where they focus on new methods that are demonstrated to be of general applicability and importance to structural biology. Articles describing particularly difficult problems in structural biology are also welcomed, if the analysis would provide useful insights to others facing similar problems.
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