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首页> 外文期刊>Infection and immunity >Identification of D motif epitopes in Staphylococcus aureus fibronectin-binding protein for the production of antibody inhibitors of fibronectin binding.
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Identification of D motif epitopes in Staphylococcus aureus fibronectin-binding protein for the production of antibody inhibitors of fibronectin binding.

机译：鉴定金黄色葡萄球菌纤连蛋白结合蛋白中的D基元表位，以产生纤连蛋白结合的抗体抑制剂。

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A fibronectin-binding protein (FnBP) adhesin of Staphylococcus aureus possesses three 37- or 38-amino-acid motifs (D1, D2, and D3) that can each bind fibronectin (Fn) with low affinity and that in tandem comprise D1-3, a high-affinity Fn-binding domain. To identify epitopes for the generation of adhesion-blocking antibodies, rabbits were immunized with recombinant D1-3 or with a glutathione S-transferase fusion protein, GSTD1-3. Affinity-purified antibodies from the D1-3 immunization were poor inhibitors of Fn binding to S. aureus and recognized several different epitopes, with a preference for clusters of acidic amino acids that do not contribute to Fn binding. Antibodies generated with GSTD1-3 as an immunogen were more effective inhibitors, but concentrations in excess of 20 microg x ml-1 did not promote more than 50% inhibition. These antibodies were highly specific for amino acids 21 to 34 of D1 (D1(21-34)), which contain a sequence that is essential for Fn binding and are identical to D2 at 12 of 14 residues. Neither antibody preparation recognized D3(20-33) of the D3 motif, where the only homology to D1(21-34) and D2(21-34) comprises a sequence motif, GG(X3,4)(I/V)DF, that is critical to Fn binding. However, antibodies specific for both D1(21-34) and D3(20-33) could be obtained by using synthetic peptides corresponding to these sequences as immunogens. F(ab')2 fragments derived from these antibodies each caused 40 to 50% inhibition of Fn binding to S. aureus, and their ability to bind to purified FnBP was eliminated by competing Fn. However, mixtures of the two F(ab')2 preparations did not provide additive or synergistic inhibition of Fn binding. Therefore, inhibition of Fn binding to S. aureus requires antibodies specific for D1(21-34) and D3(20-33), but a mixture of antibodies specific for both sequences did not provide complete inhibition.
机译：金黄色葡萄球菌的纤连蛋白结合蛋白（FnBP）粘附素具有三个37个或38个氨基酸的基序（D1，D2和D3），每个基序可以低亲和力结合纤连蛋白（Fn），并且串联构成D1-3 ，一个高亲和力的Fn结合域。为了鉴定用于产生粘附阻断抗体的表位，用重组D1-3或谷胱甘肽S-转移酶融合蛋白GSTD1-3免疫兔。来自D1-3免疫的亲和纯化抗体是Fn与金黄色葡萄球菌结合的弱抑制剂，并识别了几种不同的表位，偏爱不有助于Fn结合的酸性氨基酸簇。以GSTD1-3作为免疫原产生的抗体是更有效的抑制剂，但浓度超过20 microg x ml-1不会产生超过50％的抑制作用。这些抗体对D1（D1（21-34））的21至34位氨基酸具有高度特异性，该氨基酸包含Fn结合必不可少的序列，并且与14个残基中的12个D2相同。两种抗体制剂均不能识别D3基序的D3（20-33），其中与D1（21-34）和D2（21-34）的唯一同源性包括序列基序GG（X3,4）（I / V）DF ，这对于Fn绑定至关重要。但是，可以通过使用对应于这些序列的合成肽作为免疫原来获得对D1（21-34）和D3（20-33）都特异的抗体。源自这些抗体的F（ab'）2片段各自引起Fn与金黄色葡萄球菌结合的抑制40％至50％，并且它们与纯化的FnBP结合的能力被竞争性Fn消除。但是，两种F（ab'）2制剂的混合物未提供Fn结合的累加或协同抑制作用。因此，抑制Fn与金黄色葡萄球菌结合需要D1（21-34）和D3（20-33）特异的抗体，但对这两个序列特异的抗体混合物不能提供完全的抑制作用。

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