Reina Jose, Lacroix Emmanuel, Hobson Scott D, Fernandez-Ballester Gregorio, Rybin Vladimir, Schwab Markus S, Serrano Luis, Gonzalez Cayetano
Cell Biology and Biophysics Program, EMBL, 69117 Heidelberg, Germany.
Nat Struct Biol. 2002 Aug;9(8):621-7. doi: 10.1038/nsb815.
PDZ domains are small globular domains that recognize the last 4-7 amino acids at the C-terminus of target proteins. The specificity of the PDZ-ligand recognition is due to side chain-side chain interactions, as well as the positioning of an alpha-helix involved in ligand binding. We have used computer-aided protein design to produce mutant versions of a Class I PDZ domain that bind to novel Class I and Class II target sequences both in vitro and in vivo, thus providing an alternative to primary antibodies in western blotting, affinity chromatography and pull-down experiments. Our results suggest that by combining different backbone templates with computer-aided protein design, PDZ domains could be engineered to specifically recognize a large number of proteins.
PDZ结构域是一种小的球状结构域，可识别靶蛋白C末端的最后4至7个氨基酸。PDZ配体识别的特异性归因于侧链-侧链相互作用，以及参与配体结合的α-螺旋的定位。我们利用计算机辅助蛋白质设计制备了I类PDZ结构域的突变体，该突变体在体外和体内均可与新的I类和II类靶序列结合，从而在蛋白质印迹、亲和色谱和下拉实验中提供了一种替代一抗的方法。我们的结果表明，通过将不同的主链模板与计算机辅助蛋白质设计相结合，可以对PDZ结构域进行工程改造，使其特异性识别大量蛋白质。
