Molecular characterization of GroES and GroEL homologues from Clostridium botulinum
Molecular characterization of GroES and GroEL homologues from Clostridium botulinum
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DOI：
10.1023/a:1023028113566
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发表时间：
2003-01-01
期刊：
影响因子：
--
通讯作者：
Ohyama, T
中科院分区：
文献类型：
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作者：
Sagane, Y;Hasegawa, K;Ohyama, T
我们报告了在 D 型肉毒梭菌菌株 4947 (D-4947) 培养物上清液中 SDS-PAGE 上发现的大量 60-和 10-kDa 蛋白质的新发现。纯化蛋白的N端氨基酸序列与其他细菌GroEL和GroES蛋白的N端氨基酸序列密切相关，并且均与大肠杆菌GroEL和GroES抗体发生阳性交叉反应。作为寡聚复合物的天然 GroEL 同源物是一种弱 ATP 酶，其活性因 GroES 同源物的存在而受到抑制。通过 PCR 分离 D-4947 的 2634 bp groESL 操纵子并测序。该序列包含两个完整的开放阅读框（282和1629 bp），与细菌蛋白的groES和groEL基因家族同源。 Southern 和 Northern 印迹分析表明 groESL 操纵子作为单个拷贝在 D-4947 的基因组 DNA 上编码，而不是在其特定的毒素转化噬菌体的基因组 DNA 上编码。
We report novel findings of significant amounts of 60- and 10-kDa proteins on SDS-PAGE in a culture supernatant of the Clostridium botulinum type D strain 4947 (D-4947). The N-terminal amino acid sequences of the purified proteins were closely related to those of other bacterial GroEL and GroES proteins, and both positively cross-reacted with Escherichia coli GroEL and GroES antibodies. Native GroEL homologue as an oligomeric complex is a weak ATPase whose activity is inhibited by the presence of GroES homologue. The 2634-bp groESL operon of D-4947 was isolated by PCR and sequenced. The sequence included two complete open reading frames ( 282 and 1629 bp), which were homologous to the groES and groEL gene family of bacterial proteins. Southern and Northern blot analyses indicate that the groESL operon is encoded on the genomic DNA of D-4947 as a single copy, and not on that of its specific toxin-converting phage.