在大肠杆菌中产生的重组人白细胞介素-2的纯化及特性鉴定

Purification and characterization of recombinant human interleukin-2 produced in Escherichia coli.

作者信息

Kato K, Yamada T, Kawahara K, Onda H, Asano T, Sugino H, Kakinuma A

出版信息

Biochem Biophys Res Commun. 1985 Jul 31;130(2):692-9. doi: 10.1016/0006-291x(85)90472-3.

Abstract

Recombinant human interleukin-2 (rIL-2) produced in Escherichia coli was purified to apparent homogeneity by cation exchange chromatography and reverse phase high performance liquid chromatography. The amino acid composition, amino terminal amino acid sequence, and carboxyl terminal amino acid were consistent with those deduced from the cDNA sequence. Besides the molecular species with the amino terminal Ala, the purified preparation contained another species having an additional Met residue at the amino terminus corresponding to the initiation codon AUG. The molar absorption coefficient of rIL-2 was determined to be 9.58 X 10(3) M-1 cm-1 at 280nm in water. Ultracentrifugal analyses revealed that it existed as a monomeric form in 0.1 M NaCl. The apparent sedimentation coefficient (S20,w) was calculated to be 1.8 S.

摘要

在大肠杆菌中产生的重组人白细胞介素-2（rIL-2）通过阳离子交换色谱和反相高效液相色谱纯化至表观均一。其氨基酸组成、氨基末端氨基酸序列和羧基末端氨基酸与从cDNA序列推导的一致。除了氨基末端为丙氨酸的分子种类外，纯化制剂还包含另一种在氨基末端对应起始密码子AUG处有额外甲硫氨酸残基的种类。rIL-2在水中280nm处的摩尔吸收系数测定为9.58×10³ M⁻¹ cm⁻¹。超速离心分析表明，它在0.1 M NaCl中以单体形式存在。表观沉降系数（S₂₀,w）计算为1.8 S。