Pseudomonas aeruginosa binds to neoglycoconjugates bearing mucin carbohydrate determinants and predominantly to sialyl-Lewis x conjugates.

Pseudomonas aeruginosa binds to neoglycoconjugates bearing mucin carbohydrate determinants and predominantly to sialyl-Lewis x conjugates.
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铜绿假单胞菌与带有粘蛋白碳水化合物决定簇的新糖缀合物结合,并且主要与唾液酸-刘易斯 x 缀合物结合。

DOI:
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发表时间:
1999
期刊:
影响因子:
4.3
通讯作者:
Joël Mazurier
Joël Mazurier
中科院分区:
生物学3区
文献类型:
--
作者:
A. Scharfman;S. Degroote;Julie Beau;G. Lamblin;Philippe Roussel;Joël Mazurier

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铜绿假单胞菌在患有囊性纤维化的患者的气道的定殖中起重要作用。它与呼吸道和唾液粘蛋白的碳水化合物部分结合,并且其与囊性纤维化粘蛋白的结合甚至更高,这表明碳水化合物链的定性或/和定量修饰可能参与该过程。为了找到铜绿假单胞菌的最佳碳水化合物受体,开发了使用荧光素标记的一组基于聚丙烯酰胺的糖缀合物的流式细胞术技术。新糖缀合物含有中性、唾液酸化或硫酸化链,类似于在呼吸道粘蛋白周围发现的碳水化合物决定簇(Le(a)、Le(y)、Le(x)、唾液酸化-和3 '-磺基-Le(x)以及血型A决定簇)。我们还使用了含有半乳糖(α 1 -2)半乳糖β和唾液酸- N -乙酰乳糖胺决定簇的新糖缀合物.除了含有血型A或唾液酸- N -乙酰乳糖胺表位的糖缀合物外,这些糖缀合物与铜绿假单胞菌的无纤毛菌株1244-NP的相互作用是饱和的. Kd的测量表明,菌株1244-NP对带有唾液酸-Le(x)决定簇的糖缀合物的亲和力高于对所有其他所研究的糖缀合物的亲和力。唾液酸的作用通过主要使用唾液酸化粘蛋白糖肽的竞争测定来证实。为了发现病理菌株的这种行为是否与1244-NP突变体相同,用Le(x)新糖缀合物、其唾液酸化衍生物和其硫酸化衍生物分析了从囊性纤维化患者分离的铜绿假单胞菌的四种粘液样菌株。观察到这些菌株与三种糖缀合物结合的个体变化。然而,四个菌株中的三个菌株对唾液酸-Le(x)的亲和力高于对3 '-磺基-Le(x)衍生物的亲和力。
Pseudomonas aeruginosa plays an important role in the colonization of the airways of patients suffering from cystic fibrosis. It binds to the carbohydrate part of respiratory and salivary mucins and its binding to cystic fibrosis mucins is even higher, suggesting that qualitative or/and quantitative modifications of the carbohydrate chains may be involved in this process. In order to find out the best carbohydrate receptors for P.aeruginosa, a flow cytometry technique using a panel of polyacrylamide based glycoconjugates labeled with fluorescein was developed. The neoglycoconjugates contained neutral, sialylated or sulfated chains analogous to carbohydrate determinants found at the periphery of respiratory mucins (Le(a), Le(y), Le(x), sialyl- and 3'-sulfo-Le(x), and blood group A determinants). We used also neoglycoconjugates containing Gal(alpha1-2)Galbeta and sialyl- N -acetyllactosamine determinants. The interaction of these glycoconjugates with the nonpiliated strain of P.aeruginosa, 1244-NP, was saturable except for the glycoconjugates containing blood group A or sialyl- N -acetyllactosamine epitopes. The measure of Kd indicated that strain 1244-NP had a higher affinity for the glycoconjugate bearing the sialyl-Le(x)determinant than for all the other glycoconjugates studied. The role of sialic acid was confirmed by competition assay using mainly sialylated mucin glycopeptides. In order to find out if this behavior was the same for pathological strains as for the 1244-NP mutant, four mucoid strains of P.aeruginosa isolated from cystic fibrosis patients were analyzed with the Le(x)neoglycoconjugate, its sialylated and its sulfated derivatives. Individual variations in the binding of these strains to the three glycoconjugates were observed. However, three strains out of four had a higher affinity for the sialyl-Le(x)than for the 3'-sulfo-Le(x)derivative.
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