Freund C, Ross A, Guth B, Plückthun A, Holak T A
Department of Structural Research, Max-Planck-Institut für Biochemie, Martinsried, Germany.
FEBS Lett. 1993 Apr 5;320(2):97-100. doi: 10.1016/0014-5793(93)80070-b.
A comparison of the single-chain Fv fragment of the antibody McPC603 (scFv) with its corresponding unlinked Fv fragment has been carried out with 15N-edited NMR spectroscopy. The two Fv fragments adopt the same structure, indicating that the linker does not perturb the folding of the domains. This also directly demonstrates that folding in vivo (Fv fragment) and in vitro (scFv fragment) leads to the same structure. The main differences in the spectra of the uniformly 15N-labeled scFv and Fv fragments are due to signals of Gly and Ser from the linker peptide of the scFv fragment. The linker peptide has been mapped with NMR spectra of 15N-glycine- and 15N-glycine/15N-serine-labeled scFv fragments. The 15N T2 relaxation data indicate that the linker peptide is more flexible than the rest of the molecule.
利用15N编辑的核磁共振光谱对抗体McPC603的单链Fv片段（scFv）及其相应的非连接Fv片段进行了比较。这两个Fv片段具有相同的结构，表明连接子不会干扰结构域的折叠。这也直接证明了体内（Fv片段）和体外（scFv片段）折叠会导致相同的结构。均匀15N标记的scFv和Fv片段光谱的主要差异源于scFv片段连接肽中甘氨酸和丝氨酸的信号。通过15N-甘氨酸和15N-甘氨酸/15N-丝氨酸标记的scFv片段的核磁共振光谱对连接肽进行了定位。15N T2弛豫数据表明连接肽比分子的其余部分更具柔性。
