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烟曲霉菌丝体表面肽聚糖甘露聚糖的分析

Analysis of peptidogalactomannans from the mycelial surface of Aspergillus fumigatus.

作者信息

Haido R M, Silva M H, Ejzemberg R, Leitão E A, Hearn V M, Evans E G, Barreto Bergter E

机构信息

Instituto Biomédico, Universidade do Rio de Janeiro, Brasil.

出版信息

Med Mycol. 1998 Oct;36(5):313-21.

Abstract

Peptidogalactomannans (pGMs) from mycelium of two strains of Aspergillus fumigatus were fractionated by Cetavlon precipitation and size exclusion chromatography and their carbohydrate structures analysed using methylation-fragmentation analysis, partial acetolysis and 13C-nuclear magnetic resonance spectroscopy. The most significant difference between the pGMs of the two strains was the degree of branching and the proportion of non-reducing ends of alpha-D-Manp and beta-D-Galf units. Methylation data showed that the pGM from AF 2109 contained alpha-D-Manp and beta-D-Galf non-reducing end units in a proportion of 3:1 while, in contrast, the proportion of these structures in pGM from AF 2140 was 7:1, resulting in a highly branched structure. The immunoreactivity of the pGM fractions was tested by indirect immunofluorescence. The fractions were also tested in an ELISA system with rabbit antiserum raised to whole cells of A. fumigatus NCPF 2140 and with serum from patients with either proven aspergilloma or ABPA. The carbohydrate moiety of the pGM appears to be responsible for the antigenicity. Periodate treatment, partial acid hydrolysis and beta-elimination removed most of the antibody binding capacity.

摘要

对两株烟曲霉菌丝体中的肽聚糖半乳甘露聚糖(pGMs)进行十六烷基三甲基溴化铵沉淀和尺寸排阻色谱分离,并使用甲基化碎片化分析、部分乙酰解和13C核磁共振光谱对其碳水化合物结构进行分析。两株菌的pGMs之间最显著的差异在于α-D-甘露糖(α-D-Manp)和β-D-半乳糖(β-D-Galf)单元的分支程度和非还原端比例。甲基化数据表明,AF 2109的pGM中α-D-Manp和β-D-Galf非还原端单元的比例为3:1,而相比之下,AF 2140的pGM中这些结构的比例为7:1,形成了高度分支的结构。通过间接免疫荧光测试pGM组分的免疫反应性。这些组分还在酶联免疫吸附测定(ELISA)系统中进行测试,该系统使用针对烟曲霉NCPF 2140全细胞产生的兔抗血清以及来自确诊曲菌球或变应性支气管肺曲霉病(ABPA)患者的血清。pGM的碳水化合物部分似乎是抗原性的原因。高碘酸盐处理、部分酸水解和β-消除去除了大部分抗体结合能力。

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