Akey C W, Szalay M, Edelstein S J
Ultramicroscopy. 1984;13(1-2):103-11. doi: 10.1016/0304-3991(84)90061-5.
The molecular packing of a trigonal crystal form of catalase initially studied by Longley [1] has been re-evaluated. Sections of crystals fixed and preserved with tannic acid were obtained parallel to the (001) and (100) planes. Specimens prepared by either conventional or low temperature embedding maintained 20 A resolution after sectioning. The space group of the crystals is either P3(1)21 or P3(2)21 and the observed unit cell parameters for (001) and (100) are a = b = 174 A, gamma = 119 degrees and b = 189 A, c = 248 A with alpha = 89.5 degrees. Computer-based reconstructions of two principal projections coupled with crystal density measurements allowed the deduction that there is one catalase tetramer per asymmetric unit. The crystal structure consists of 6 molecules packed closely about a common triad screw axis. This interpretation differs from that proposed by Longley [J. Mol. Biol. 30 (1967) 323], because thin sections of embedded crystals were assumed a priori to be positively stained in the early work; in actuality the sections were negatively stained. We also demonstrate that tannic acid fixation can lead to well preserved, positively stained crystal sections under certain conditions.
最初由朗利[1]研究的过氧化氢酶三角晶体形式的分子堆积情况已被重新评估。获得了用单宁酸固定和保存的与(001)和(100)平面平行的晶体切片。通过常规或低温包埋制备的标本在切片后保持了20埃的分辨率。晶体的空间群为P3(1)21或P3(2)21,观察到的(001)和(100)的晶胞参数为a = b = 174埃,γ = 119度,b = 189埃,c = 248埃,α = 89.5度。基于计算机的两个主投影重建以及晶体密度测量使得能够推断出每个不对称单元中有一个过氧化氢酶四聚体。晶体结构由围绕一个共同的三重螺旋轴紧密堆积的6个分子组成。这种解释与朗利[《分子生物学杂志》30(1967)323]提出的不同,因为在早期工作中,先验地假设包埋晶体的薄片是正染色的;实际上这些切片是负染色的。我们还证明,在某些条件下,单宁酸固定可以导致保存良好的正染色晶体切片。