Manjula B N, Mushinski E B, Glaudemans C P
J Immunol. 1977 Sep;119(3):867-71.
Murine myeloma immunoglobulin (IgA, K) J539, which shows enhanced tryptophanyl fluorescence on ligand binding, and S10, which shows reverse-sign changes in tryptophanyl fluorescence on ligand binding (RLIF, see below), have been reduced, alkylated, and dissociated into their light (L) and heavy (H) chains. Two hybrid recombinants, H10L539 and H539L10, have been prepared and the 7S material has been isolated by chromatography. The binding behavior of these recombinants was studied with a number of ligands. Both recombinants showed activity with beta(1 leads to 6) linked galactose ligands comparable to the native immunoglobulins. The ligand-induced fluorescence changes of the recombinants paralleled those of the heavy chain donor. For the recombinant H10L539, two different galactose-ligands caused fluorescence changes in opposite directions. It was quantitatively shown that binding of these ligands, nevertheless, took place in the same combining region. The idiotype of each recombinant resembled that of the heavy chain donor.
小鼠骨髓瘤免疫球蛋白(IgA,κ)J539在与配体结合时色氨酸荧光增强,而S10在与配体结合时色氨酸荧光呈现反向信号变化(RLIF,见下文),已对其进行还原、烷基化处理,并解离为轻链(L)和重链(H)。制备了两种杂交重组体H10L539和H539L10,并通过色谱法分离出了7S物质。用多种配体研究了这些重组体的结合行为。两种重组体对β(1→6)连接的半乳糖配体均表现出与天然免疫球蛋白相当的活性。重组体的配体诱导荧光变化与重链供体的荧光变化相似。对于重组体H10L539,两种不同的半乳糖配体引起的荧光变化方向相反。但定量结果表明,这些配体的结合发生在相同的结合区域。每个重组体的独特型与重链供体的相似。