Müller M, Affolter M, Leupin W, Otting G, Wüthrich K, Gehring W J
Biozentrum der Universität Basel, Switzerland.
EMBO J. 1988 Dec 20;7(13):4299-304. doi: 10.1002/j.1460-2075.1988.tb03328.x.
The homeodomain encoded by the Antennapedia (Antp) gene of Drosophila was overproduced in a T7 expression vector in Escherichia coli. The corresponding polypeptide of 68 amino acids was purified to homogeneity. The homeodomain was analysed by ultracentrifugation and assayed for DNA binding. The secondary structure of the isolated homeodomain was determined by nuclear magnetic resonance spectroscopy. DNA-binding studies indicate that the isolated homeodomain binds to DNA in vitro. It selectively binds to the same sites as a longer Antp polypeptide and a full-length fushi tarazu (ftz) protein. Therefore, the homeodomain represents the DNA-binding domain of the homeotic proteins.
果蝇触角足(Antp)基因编码的同源异型结构域在大肠杆菌的T7表达载体中过量表达。由68个氨基酸组成的相应多肽被纯化至同质。通过超速离心分析该同源异型结构域,并检测其与DNA的结合能力。通过核磁共振光谱法确定分离出的同源异型结构域的二级结构。DNA结合研究表明,分离出的同源异型结构域在体外能与DNA结合。它与较长的Antp多肽和全长的分节基因(ftz)蛋白选择性地结合于相同位点。因此,同源异型结构域代表了同源异型蛋白的DNA结合结构域。