Rauhut R, Gabius H J, Kühn W, Cramer F
J Biol Chem. 1984 May 25;259(10):6340-5.
Phenylalanyl-tRNA synthetase from the archaebacterium Methanosarcina barkeri was purified 1620-fold with 24% overall yield. It appears to be a tetrameric enzyme with a molecular mass of 270 kDa, as determined by gel filtration, with a subunit structure of alpha 2 beta 2 (alpha = 63 kDa, beta = 70 kDa), as determined by sodium dodecyl sulfate gel electrophoresis. No conservation of common antigenic determinants is noted with polyclonal antibodies raised against the enzymes of Escherichia coli, yeast, and hen liver. Heterologous aminoacylation of tRNA with high selectivity for archaebacterial tRNA and substrate properties of ATP analogues reveals a unique pattern, reflecting the supposed genealogical difference between the urkingdoms of archaebacteria, eubacteria, and eukaryotes.
从嗜甲烷八叠球菌中纯化得到的苯丙氨酰 - tRNA合成酶,纯化倍数达1620倍,总产率为24%。通过凝胶过滤测定,它似乎是一种分子量为270 kDa的四聚体酶,而通过十二烷基硫酸钠凝胶电泳测定,其亚基结构为α2β2(α = 63 kDa,β = 70 kDa)。用针对大肠杆菌、酵母和鸡肝的酶产生的多克隆抗体未发现共同抗原决定簇的保守性。对古细菌tRNA具有高选择性的tRNA异源氨酰化以及ATP类似物的底物特性揭示了一种独特模式,反映了古细菌、真细菌和真核生物这三个超界之间推测的谱系差异。