Nonaromatic amino acids in the combining site region of a monoclonal anti-spin-label antibody.
Nonaromatic amino acids in the combining site region of a monoclonal anti-spin-label antibody.
复制标题
单克隆抗自旋标记抗体的结合位点区域中的非芳香族氨基酸。
DOI：
10.1021/bi00321a030
复制
发表时间：
1984
期刊：
影响因子：
2.9
通讯作者：
McConnell,HM
中科院分区：
文献类型：
--
作者：
Frey,T;Anglister,J;McConnell,HM
材料和方法已经描述了自旋标记半抗原的合成（Balakrishnan等人，1982年）。化学式也可以在Anglister et al.（1984年a）。AN 02细胞系的起源、维持和标记已经在先前描述（Anglister等人，1984年a）。通过标准程序制备Fab片段。全氘代氨基酸L-赖氨酸、L-苏氨酸和L-异亮氨酸购自剑桥同位素实验室。全氘代L-丙氨酸、L-亮氨酸、L-缬氨酸和1-[甲基-2H3]甲硫氨酸购自MSD Isotopes。用氘代丙氨酸标记的培养基包括3-4 mM丙氨酸。在所有其他情况下，RPMI培养基中的氨基酸量正常。用JEOL 500-MHz光谱仪获取NMR光谱。Fab的浓度范围为（1.3-3）× 10~ 4 M，进样量为550 μ L。在8000个数据点中收集自由诱导衰变，
Materials and MethodsThe synthesis of the spin-label hapten has been described (Balakrishnan et al., 1982). The chemicalformula can also be found in Anglister et al.(1984a). The origin, maintenance, and labeling of the AN02 cell line have been described previously (Anglister et al., 1984a). Fab fragments were prepared by standard procedures. The amino acids perdeuterated L-lysine, L-threonine, and L-isoleucine were purchased from Cambridge Isotopes Laboratories. Per-deuterated L-alanine, L-leucine, L-valine, and l-[methyl-2H3] methionine were purchased from MSD Isotopes. Medium labeled with deuterated alanine included 3-4 mM alanine. In all other cases, the amino acids were in the regular quantities for RPMI medium. NMR spectra were taken with a JEOL 500-MHz spectrometer. The concentration of Fab was in the range (1.3-3) X 10~ 4 M, and sample volumes were 550¡ uL. Free induction decays were collected in 8000 data points after