Habeeb A F
Biochim Biophys Acta. 1975 Nov 18;412(1):62-9. doi: 10.1016/0005-2795(75)90339-6.
epsilon-Prototoxin of clostridium perfringens type D consists of one polypeptide chain of 311 amino acids with the following composition: Asp52 Thr31 Ser25 Glu28 Pro12 Gly17 Ala14 Val28 Met5 Ile15 Leu18 Tyr17 Phe8 Lys31 His2 Arg5 Tyr2. It has no free cysteine but contains one blocked cysteine. The N-terminal as well as the C-terminal residue is lysine. The ultracentrifuge pattern gave one single boundary having S020,w = 2.15 S and Do20,w = 5.56-10(-7) cm2/s. Calculation of the molecular weight from D020,w and S020,w gave a value of 34 250. The molecular weight determined from sedimentation equilibrium using ultraviolet optics gave a value of 33 000 +/- 1000. On the other hand molecular weights calculated from a calibrated Sephadex G-100 column in borate buffer was 25 000 and that in sodium phosphate, ionic strength 0.2, was 27 500. This discrepancy between values obtained in the ultracentrifuge and by gel filtration is attributed to adsorption of epsilon-prototoxin by Sephadex.
D型产气荚膜梭菌的ε-原毒素由一条含311个氨基酸的多肽链组成,其氨基酸组成如下:天冬氨酸52个、苏氨酸31个、丝氨酸25个、谷氨酸28个、脯氨酸12个、甘氨酸17个、丙氨酸14个、缬氨酸28个、甲硫氨酸5个、异亮氨酸15个、亮氨酸18个、酪氨酸17个、苯丙氨酸8个、赖氨酸31个、组氨酸2个、精氨酸5个、酪氨酸2个。它不含游离半胱氨酸,但含有一个封闭的半胱氨酸。N末端和C末端残基均为赖氨酸。超速离心图谱给出一个单一的边界,S020,w = 2.15 S,D020,w = 5.56×10(-7) cm2/s。根据D020,w和S020,w计算的分子量为34250。使用紫外光学通过沉降平衡测定的分子量为33000±1000。另一方面,在硼酸盐缓冲液中通过校准的Sephadex G - 100柱计算的分子量为25000,在离子强度为0.2的磷酸钠中为27500。超速离心和凝胶过滤得到的值之间的这种差异归因于Sephadex对ε-原毒素的吸附。