Stroud R M, Serwer P, Ross M J
Biophys J. 1981 Dec;36(3):743-57. doi: 10.1016/S0006-3495(81)84763-7.
The dimensions of bacteriophage T7 and T7 capsids have been investigated by small-angle x-ray scattering. Phage T7 behaves like a sphere of uniform density with an outer radius of 301 +/- 2 A (excluding the phage tail) and a calculated volume for protein plus nucleic acid of 1.14 +/- 0.05 x 10(-16) ml. The outer radius determined for T7 phage in solution is approximately 30% greater than the radius measured from electron micrographs, which indicates that considerable shrinkage occurs during preparation for electron microscopy. Capsids that have a phagelike envelope and do not contain DNA were obtained from lysates of T7-infected Escherichia coli (capsid II) and by separating the capsid component of T7 phage from the phage DNA by means of temperature shock (capsid IV). In both cases the peak protein density is at a radius of 275 A; the outer radius is 286 +/- 4 A, approximately 5% smaller than the envelope of T7 phage. The thickness of the envelope of capsid II is 22 +/- 4 A, consistent with the thickness of protein estimated to be 23 +/- 5 A in whole T7 phage, as seen on electron micrographs in which the internal DNA is positively stained. The volume in T7 phage available to package DNA is estimated to be 9.2 +/- 0.4 x 10(-17) ml. The packaged DNA adopts a regular packing with 23.6 A interplanar spacing between, DNA strands. The angular width of the 23.6 A reflection shows that the mean DNA-DNA spacing throughout the phage head is 27.5 +/- less than 2.2 A. A T7 precursor capsid (capsid I) expands when pelleted for x-ray scattering in the ultracentrifuge to essentially the same outer dimensions as for capsids II and IV. This expansion of capsid I can be prevented by fixing with glutaraldehyde; fixed capsid I has peak density at a radius of 247 A, 10% less than capsid II or IV.
利用小角X射线散射研究了噬菌体T7及其衣壳的尺寸。噬菌体T7表现得像一个密度均匀的球体,其外半径为301±2埃(不包括噬菌体尾部),蛋白质加核酸的计算体积为1.14±0.05×10⁻¹⁶毫升。在溶液中测定的T7噬菌体的外半径比从电子显微镜照片测得的半径大约大30%,这表明在制备电子显微镜样品的过程中发生了相当大的收缩。具有噬菌样包膜且不含DNA的衣壳是从T7感染的大肠杆菌裂解物中获得的(衣壳II),也可通过温度休克将T7噬菌体的衣壳成分与噬菌体DNA分离得到(衣壳IV)。在这两种情况下,蛋白质密度峰值位于半径275埃处;外半径为286±4埃,比T7噬菌体的包膜小约5%。衣壳II包膜的厚度为22±4埃,与在电子显微镜照片中内部DNA被阳性染色时,整个T7噬菌体中估计的蛋白质厚度23±5埃一致。估计T7噬菌体中可用于包装DNA的体积为9.2±0.4×10⁻¹⁷毫升。包装的DNA采用规则堆积,DNA链之间的面间距为23.6埃。23.6埃反射的角宽度表明,整个噬菌体头部的平均DNA-DNA间距为27.5±小于2.2埃。当T7前体衣壳(衣壳I)在超速离心机中沉淀以进行X射线散射时,它会膨胀到与衣壳II和IV基本相同的外部尺寸。用戊二醛固定可以防止衣壳I的这种膨胀;固定后的衣壳I在半径247埃处有密度峰值,比衣壳II或IV小10%。