Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT 06268.
Proc Natl Acad Sci U S A. 1987 Sep;84(17):6126-30. doi: 10.1073/pnas.84.17.6126.
Frankia sp. strain CpI1 has two glutamine synthetases. Glutamine synthetase I (GSI) is present during growth on ammonium or N(2) and is similar to classical prokaryotic glutamine synthetases. Gel-filtration chromatography gave a molecular weight estimate of about 680,000 for the GSI holoenzyme, and denaturing polyacrylamide gel electrophoresis yielded a subunit molecular weight of about 59,000, indicating that GSI is most likely a dodecamer. GSI is regulated by adenylylation, as shown by the presence of two spots on two-dimensional polyacrylamide gel electrophoresis and by its behavior during treatment with snake venom phosphodiesterase. GSII is derepressed during nitrogen starvation and accounts for about 95% of the glutamine synthetase activity in nitrogen-starved cells. It is heat-labile and has a subunit molecular weight of about 43,000. Frankia GSII is similar to GSII enzymes found in all but one member of the Rhizobiaceae analyzed to date. The presence of a second glutamine synthetase in Frankia lends support to the proposal that symbiotic organisms have unique modes of nitrogen nutrition but reopens questions about the origins and uniqueness of GSII genes in members of the Rhizobiaceae.
弗兰克氏菌菌株 CpI1 有两种谷氨酰胺合成酶。谷氨酰胺合成酶 I(GSI)在铵或 N2 上生长时存在,与经典的原核谷氨酰胺合成酶相似。凝胶过滤层析法估计 GSI 全酶的分子量约为 680,000,变性聚丙烯酰胺凝胶电泳得到的亚基分子量约为 59,000,表明 GSI 很可能是十二聚体。GSI 通过腺苷酰化调节,如二维聚丙烯酰胺凝胶电泳上存在两个斑点和用蛇毒磷酸二酯酶处理时的行为所示。GSII 在氮饥饿时被解除阻遏,占氮饥饿细胞中谷氨酰胺合成酶活性的约 95%。它是热不稳定的,亚基分子量约为 43,000。弗兰克氏菌 GSII 与迄今为止分析的根瘤菌科除一个成员外的所有 GSII 酶相似。弗兰克氏菌中存在第二种谷氨酰胺合成酶支持共生生物具有独特的氮营养模式的观点,但重新引发了关于根瘤菌科成员中 GSII 基因的起源和独特性的问题。