Gerard N P, Lively M O, Gerard C
Department of Medicine, Beth Israel Hospital, Boston, MA 02215.
Protein Seq Data Anal. 1988;1(6):473-8.
The anaphylatoxin peptide, C3a, was isolated in its inactive des-arginine form from complement-activated guinea pig serum and the complete amino acid sequence was determined. The peptide was isolated using a modification of previously described methods for anaphylatoxin purification and yielded 18 mg of guinea C3ades Arg judged homogeneous by cellulose acetate electrophoresis, high performance liquid chromatography, amino acid composition, and sequence analyses. The complete amino acid sequence of the molecule was determined by automated Edman degradation of intact C3ades Arg following performic acid oxidation, and of four peptides obtained from CNBr, endoproteinase Lys-C, or endoproteinase Glu-C (V8 protease) digests of reduced and alkylated C3ades Arg. The sequence is as follows: Guinea pig C3ades Arg has approximately 70% overall sequence identity with human, porcine, rat and mouse anaphylatoxins. As with C3a's from other species, the guinea pig molecule has an invariant C-terminal tetrapeptide sequence, LGLA, and six invariant cysteinyl residues at positions 23, 24, 37, 50, 57, and 58.
过敏毒素肽C3a以其无活性的去精氨酸形式从补体激活的豚鼠血清中分离出来,并测定了其完整的氨基酸序列。该肽是采用对先前描述的过敏毒素纯化方法的一种改进而分离得到的,得到了18mg经醋酸纤维素电泳、高效液相色谱、氨基酸组成和序列分析判断为纯一的豚鼠C3ades Arg。该分子的完整氨基酸序列是通过对完整的C3ades Arg进行过甲酸氧化后的自动Edman降解,以及对还原和烷基化的C3ades Arg经溴化氰、内肽酶Lys-C或内肽酶Glu-C(V8蛋白酶)消化得到的四个肽段进行测定的。序列如下:豚鼠C3ades Arg与人类、猪、大鼠和小鼠的过敏毒素在总体序列上约有70%的同一性。与来自其他物种的C3a一样,豚鼠分子具有不变的C末端四肽序列LGLA,以及在第23、24、37、50、57和58位的六个不变的半胱氨酸残基。