Structural and functional characterization of Pseudomonas aeruginosa CupB chaperones.

Structural and functional characterization of Pseudomonas aeruginosa CupB chaperones.
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铜绿假单胞菌 CupB 伴侣的结构和功能表征

DOI:
10.1371/journal.pone.0016583
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发表时间:
2011-01-31
期刊:
影响因子:
3.7
通讯作者:
Meng G
Meng G
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Cai X;Wang R;Filloux A;Waksman G;Meng G

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铜绿假单胞菌是一种重要的人类病原体,据估计是全世界约 10% 的医院感染的原因。铜绿假单胞菌的发病机制始于其在受损组织或医疗器械(例如导管、假体和植入心脏瓣膜等)中的定殖,并受到包括菌毛在内的多种细胞外粘附因子的促进。先前已在铜绿假单胞菌染色体上鉴定出几个含有菌毛基因的簇,并将其命名为cup。 CupB 菌毛的组装被认为是由两个伴侣 CupB2 和 CupB4 协调的。然而,由于缺乏结构和生化数据,它们的伴侣活性仍然是推测性的。在这项研究中,我们报道了铜绿假单胞菌 CupB2 的 2.5 Å 晶体结构。基于该结构,我们使用有限胰蛋白酶消化和链霉-tactin Pull-down测定进一步测试了CupB2和CupB4对CupB1(假定的主要菌毛亚基)和CupB6(假定的粘附素)的结合特异性。结构和生化数据表明 CupB2 和 CupB4 在 CupB 分泌中可能发挥不同但并非多余的作用。 CupB2可能是CupB1的伴侣,而CupB4可能是CupB4:CupB5:CupB6的伴侣,其中CupB4和CupB6的相互作用可能是通过CupB5介导的。
Pseudomonas aeruginosa, an important human pathogen, is estimated to be responsible for ∼10% of nosocomial infections worldwide. The pathogenesis of P. aeruginosa starts from its colonization in the damaged tissue or medical devices (e.g. catheters, prothesis and implanted heart valve etc.) facilitated by several extracellular adhesive factors including fimbrial pili. Several clusters containing fimbrial genes have been previously identified on the P. aeruginosa chromosome and named cup . The assembly of the CupB pili is thought to be coordinated by two chaperones, CupB2 and CupB4. However, due to the lack of structural and biochemical data, their chaperone activities remain speculative. In this study, we report the 2.5 Å crystal structure of P. aeruginosa CupB2. Based on the structure, we further tested the binding specificity of CupB2 and CupB4 towards CupB1 (the presumed major pilus subunit) and CupB6 (the putative adhesin) using limited trypsin digestion and strep-tactin pull-down assay. The structural and biochemical data suggest that CupB2 and CupB4 might play different, but not redundant, roles in CupB secretion. CupB2 is likely to be the chaperone of CupB1, and CupB4 could be the chaperone of CupB4:CupB5:CupB6, in which the interaction of CupB4 and CupB6 might be mediated via CupB5.
DOI: 10.1073/pnas.0408072101
发表时间: 2004-12-14
影响因子: 11.1
作者:
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发表时间: 2007-05-01
影响因子: 3.2
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发表时间: 2001-06-05
影响因子: 11.1
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