Riedel H D, Heinrich J, Heisig A, Choli T, Schuster H
Max-Planck-Institut für Molekulare Genetik, Berlin, Germany.
FEBS Lett. 1993 Nov 15;334(2):165-9. doi: 10.1016/0014-5793(93)81705-5.
Two antirepressor proteins, Ant1 and Ant2, of molecular weight 42 and 32 kDa, respectively, are encoded by P1 as a single open reading frame, with the smaller protein initiating at an in-frame start codon. Another open reading frame, icd, 5' upstream of and overlapping ant1 is required for ant1 expression. Using appropriate ant gene-carrying plasmids we have overproduced and purified Ant1/2 in the form of a protein complex and Ant2 as a single protein. Sequence analysis confirmed the N-terminal amino acids predicted from the DNA sequence of ant1/ant2, except that the N-terminal methionine is missing in the Ant2 protein. Under appropriate conditions the C1 repressors of phages P1 and P7 specifically co-precipitate with the Ant1/2 complex but not with Ant2 protein alone. The results suggest that the antirepressor may exert its C1-inactivating function by a direct protein-protein interaction.
两种抗阻遏蛋白,Ant1和Ant2,分子量分别为42 kDa和32 kDa,由P1编码为一个单一的开放阅读框,较小的蛋白质从一个框内起始密码子开始。另一个开放阅读框icd,位于ant1的5'上游且与ant1重叠,是ant1表达所必需的。使用合适的携带ant基因的质粒,我们已经过量表达并纯化了蛋白质复合物形式的Ant1/2和单一蛋白质形式的Ant2。序列分析证实了从ant1/ant2的DNA序列预测的N端氨基酸,但Ant2蛋白中缺少N端甲硫氨酸。在适当条件下,噬菌体P1和P7的C1阻遏物与Ant1/2复合物特异性共沉淀,但不与单独的Ant2蛋白共沉淀。结果表明,抗阻遏蛋白可能通过直接的蛋白质-蛋白质相互作用发挥其C1失活功能。