DIFFERENTIAL PROTEIN EXPRESSION BY SHIGELLA-FLEXNERI IN INTRACELLULAR AND EXTRACELLULAR ENVIRONMENTS
DIFFERENTIAL PROTEIN EXPRESSION BY SHIGELLA-FLEXNERI IN INTRACELLULAR AND EXTRACELLULAR ENVIRONMENTS
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DOI：
10.1073/pnas.87.11.4179
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发表时间：
1990-06-01
影响因子：
11.1
通讯作者：
PAYNE, SM
中科院分区：
文献类型：
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作者：
HEADLEY, VL;PAYNE, SM
志贺氏菌固有的放射性标记与[35 S]蛋氨酸无论是在细胞外或在感染的HeLa细胞单层内繁殖。观察到蛋白质的抑制和诱导的复杂模式。约97，62，58，50，25，和18千道尔顿（kDa）的蛋白质诱导从感染的单层分离的福氏志贺菌。100，85，70，64和55 kDa的蛋白质在相同的条件下被抑制，但在单独的组织培养基中标记的细胞中观察到。蛋白质表达的阶段，附着，入侵，和细胞内增殖的脉冲标记检查。58-kDa的蛋白只在入侵过程中诱导，62-和25-kDa的蛋白只在细胞内增殖诱导。转移到离子浓度和pH模拟细胞内条件和内体pH的基本培养基中，导致7-和58-kDa蛋白的诱导，并减少细胞内样介质与2-巯基乙醇导致97-，50-，和25-kDa蛋白的诱导和55-kDa蛋白的抑制。在体外和体内生长的志贺氏菌的放射免疫沉淀显示免疫原性蛋白质的差异表达。在细胞内增殖过程中，大小对应于IpaB（62 kDa）、IpaC（42 kDa）和iPad（38 kDa）的蛋白质丢失，而另一种对应于IpaA（80 kDa）的蛋白质在相同条件下被发现增加。
Shigellae were intrinsically radiolabeled with [35S]methionine either extracellularly or while multiplying within infected HeLa cell monolayers. a complex pattern of suppression and induction of proteins was observed. Proteins of approximately 97, 62, 58, 50, 25, and 18 kilodaltons (kDa) were induced in Shigella flexneri isolated from infected monolayers. Proteins of 100, 85, 70, 64, and 55 kDa were suppressed under the same conditions but were seen in cells labeled in the tissue culture medium alone. Protein expression during the stages of attachment, invasion, and intracellular multiplication was examined by pulse-labeling. The 58-kDa protein was induced only during invasion, and the 62- and 25-kDa proteins were induced only during intracellular multiplication. Shift into a minimal medium with ion concentrations and pH mimicking intracellular conditions and endosomal pH resulted in the induction of the 7- and 58-kDa proteins, and reduction of the intracellular-like medium with 2-mercaptoethanol resulted in the induction of the 97-, 50-, nd 25-kDa proteins and suppression of the 55-kDa protein. Radioimmunoprecipitations of shigellae grown in vitro and in vivo revealed differential expression of immunogenic proteins. Proteins corresponding in size to IpaB (62 kDa), IpaC (42 kDa), and IpaD (38 kDa) were lost during intracellular multiplication, whereas another protein corresponding to IpaA (80 kDa) was found to increase under the same conditions.