Hadler N M, Metzger H
Proc Natl Acad Sci U S A. 1971 Jul;68(7):1421-4. doi: 10.1073/pnas.68.7.1421.
The gammaA myeloma protein 315 from the mouse was affinity-labeled with m-nitrobenzene diazonium fluoroborate which, as shown previously, leads to selective modification of the tyrosine at position 34 in the light chains of this protein. The azotyrosine bond was reduced with dithionite to form 3-aminotyrosine. The aryl amino group of the aminotyrosine was selectively reacted with the bifunctional reagent 1,5-difluoro-2,4-dinitrobenzene. Cross-links were formed between the aminotyrosine and at least two residues-one on the same light chain and one in the Fd region of the heavy chain. Since affinity labeling of various antibodies with diazonium reagents has frequently led to azotyrosine formation, the approach described in this paper should have general applicability.
来自小鼠的γA骨髓瘤蛋白315用间硝基苯重氮氟硼酸盐进行亲和标记,如先前所示,该试剂可导致该蛋白轻链中第34位酪氨酸的选择性修饰。用连二亚硫酸盐还原偶氮酪氨酸键以形成3-氨基酪氨酸。氨基酪氨酸的芳基氨基与双功能试剂1,5-二氟-2,4-二硝基苯选择性反应。在氨基酪氨酸与至少两个残基之间形成交联——一个在同一条轻链上,另一个在重链的Fd区域。由于用重氮试剂对各种抗体进行亲和标记经常会导致偶氮酪氨酸的形成,本文所述方法应具有普遍适用性。