Jilka R L, Pestka S
Proc Natl Acad Sci U S A. 1977 Dec;74(12):5692-6. doi: 10.1073/pnas.74.12.5692.
Partially purified mRNA coding for the MOPC-315 heavy immunoglobulin alpha chain was translated in a reticulocyte lysate containing 20 labeled amino acids. Radiolabeled MOPC-315 heavy chain precursor protein, purified by preparative gel electrophoresis and immunoprecipitation, was sequenced by Edman degradation. The labeled phenylthiohydantoin amino acid obtained in each cycle was identified and quantitated by high-pressure liquid chromatography. The precursor sequence of 18 amino acids, Met-Lys-Val-Leu-Ser-Leu-Leu-Tyr-Leu-Leu-Thr-Ala-Ile-Pro-His-Ile-Met-Ser, preceded the sequence corresponding to the NH2 terminus of the mature secreted heavy chain.
编码MOPC - 315重免疫球蛋白α链的部分纯化mRNA在含有20种标记氨基酸的网织红细胞裂解物中进行翻译。通过制备性凝胶电泳和免疫沉淀纯化的放射性标记MOPC - 315重链前体蛋白,采用埃德曼降解法进行测序。通过高压液相色谱对每个循环中获得的标记苯硫代乙内酰脲氨基酸进行鉴定和定量。18个氨基酸的前体序列,即甲硫氨酸 - 赖氨酸 - 缬氨酸 - 亮氨酸 - 丝氨酸 - 亮氨酸 - 亮氨酸 - 酪氨酸 - 亮氨酸 - 亮氨酸 - 苏氨酸 - 丙氨酸 - 异亮氨酸 - 脯氨酸 - 组氨酸 - 异亮氨酸 - 甲硫氨酸 - 丝氨酸,位于与成熟分泌重链NH2末端相对应的序列之前。