Lütticken C, Krüttgen A, Möller C, Heinrich P C, Rose-John S
Institut für Biochemie der RWTH Aachen, Germany.
FEBS Lett. 1991 May 6;282(2):265-7. doi: 10.1016/0014-5793(91)80491-k.
Thirteen point mutations of human interleukin-6 at the C-terminus were constructed at the cDNA-level. Two degenerate oligonucleotide primers were used for PCR synthesis of two groups of point mutations at positions 182 and 184. The mutated cDNAs were in vitro transcribed and translated and subsequently assayed for biological activity in the B9 cell proliferation test. Our results confirm our former findings obtained with deletion mutants on the importance of the C-terminus of IL-6 for biological activity. In addition, we now present evidence for the importance of an alpha-helix at the C-terminus of IL-6 and the presence of the positive charge at position 182 for biological activity.
在cDNA水平构建了人白细胞介素-6 C末端的13个点突变。使用两组简并寡核苷酸引物通过PCR合成第182和184位的两组点突变。对突变的cDNA进行体外转录和翻译，随后在B9细胞增殖试验中测定其生物活性。我们的结果证实了我们之前用缺失突变体获得的关于IL-6 C末端对生物活性重要性的发现。此外，我们现在提供证据表明IL-6 C末端的α-螺旋以及第182位正电荷的存在对生物活性很重要。
