Immunochemical detection of dissolved proteins and their source bacteria in marine environments
Immunochemical detection of dissolved proteins and their source bacteria in marine environments
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海洋环境中溶解蛋白及其来源细菌的免疫化学检测
DOI：
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发表时间：
1997
期刊：
影响因子：
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通讯作者：
E. Tanoue
中科院分区：
文献类型：
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作者：
S. Suzuki;K. Kogure;E. Tanoue
为了进一步阐明在海水中以溶解蛋白形式存在的特定蛋白，以及这些蛋白的来源是细菌孔蛋白的发现，我们对海洋环境和培养的细菌进行了调查，以寻找两种细菌孔蛋白的同源物。制备了铜绿假单胞菌OprP孔蛋白n端抗血清和拉氏李斯特菌Omp35La孔蛋白全分子抗血清，作为Western blot检测探针。在亚北极和亚热带太平洋以及南冰洋采集的所有样本中，检测到对抗血清有反应的蛋白质。OprP和Omp35La的分子量分别为48 kDa和33 ~ 37 kDa；海水样品中检测到的蛋白质通常也具有相似的分子质量。而对不同分子质量培养细菌的溶解蛋白和外膜蛋白进行了检测。这表明，与OprP和Oinp35La不同的溶解蛋白和细菌外膜蛋白与OprP和Omp35La具有相似的抗原结构。荧光抗体染色显示：抗血清染色存在于整个水柱的天然细菌组合中。目前的观察结果有力地表明，细菌孔蛋白是溶解蛋白的主要来源。
In order to expand upon the d~scovery that specific proteins survive in seawater as dissolved protein and that the ongin of these proteins is bacterial porin, we surveyed marine environments and cultured bacteria for the presence of homologues of 2 kinds of bacterial porins. Antisera against the N-terminus of the OprP porin of Pseudornonas aeruginosa and against the whole molecule of the Omp35La porin of Listonella (Vibdo) anguillarum were prepared and used as probes in Western blot analysis. In all samples collected in the subarctic and subtropical Pacific Ocean and the Antarctic Ocean, proteins reactive to the antisera were detected. The molecular ~nasses of OprP and Omp35La are 48 and 33 to 37 kDa respectively; detected proteins In seawater samples were generally also of similar molecular mass. However, dissolved proteins as well as outer membrane proteins from cultured bacteria with different molecular masses were detected using the antisera. This indicates that dissolved proteins and bacter~al outer membrane proteins distinct from OprP and Oinp35La contain similar antigenic structures to OprP and Omp35La. Fluorescent-antibody staining revealed that bacterial cells tha.t yere stainable with antisera were present in natural bacterial assemblages throughout the entlre water column. Present observations strongly suggest that bacterial porins are a major source of dissolved proteins.