Characterization of the WW domain of human yes-associated protein and its polyproline-containing ligands
Characterization of the WW domain of human yes-associated protein and its polyproline-containing ligands
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DOI：
10.1074/jbc.272.27.17070
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发表时间：
1997-07-04
影响因子：
4.8
通讯作者：
Sudol, M
中科院分区：
文献类型：
--
作者：
Chen, HI;Einbond, A;Sudol, M
WW结构域是由38-40个半保守氨基酸组成的一个新的球状结构域，参与介导蛋白质-蛋白质相互作用。WW结构域是酵母、线虫和哺乳动物中具有不同功能的蛋白质所共有的，包括结构蛋白、调节蛋白和信号蛋白。通过筛选16天小鼠胚胎表达文库，我们鉴定了Yes激酶相关蛋白WW结构域的两个推定配体，我们将其命名为WW结构域结合蛋白1和2。这两个蛋白通过短的富含脯氨酸的基序与WW结构域相互作用，该基序具有四个连续的脯氨酸和一个酪氨酸的She共有序列。我们报道了WW结构域结合蛋白I和2的人直系同源物的cDNA克隆和表征，这些cDNA克隆编码的产物代表了功能未知的新蛋白，此外，利用荧光原位杂交技术，将WW结构域结合蛋白1和2的基因分别定位在染色体2 p12和17 q25上。此外，使用定点诱变，我们确定了Yes激酶相关蛋白WW结构域中的哪些残基对结合至关重要。最后，通过合成肽，其中四个连续的脯氨酸-酪氨酸基序和周围的五个残基的各个位置被所有可能的氨基酸残基取代，我们进一步阐明了该基序的结合要求。
We had previously identified the WW domain as a novel globular domain that is composed of 38-40 semi-conserved amino acids and is involved in mediating protein-protein interaction, The WW domain is shared by proteins of diverse functions including structural, regulatory, and signaling proteins in yeast, nematode, and mammals, Functionally it is similar to the Src homology 3 domain in that it binds polyproline ligands. By screening a 16-day mouse embryo expression library, we identified two putative ligands of the WW domain of Yes kinase-associated protein which we named WW domain-binding proteins 1 and 2, These proteins interacted with the WW domain via a short proline-rich motif with She consensus sequence of four consecutive prolines followed by a tyrosine, Herein, we report the cDNA cloning and characterization of the human orthologs of WW domain-binding proteins I and 2, The products encoded by these cDNA clones represent novel proteins with no known function, Furthermore, these proteins show no homology to each other except for a proline-rich motif, By fluorescence in situ hybridization oil human metaphase chromosomes, we mapped the human genes for WW domain-binding proteins 1 and 2 to chromosomes 2p12 and 17q25, respectively. In addition, using site-directed mutagenesis, we determined which residues in the WW domain of Yes kinase-associated protein are critical for binding. Finally, by synthesizing peptides in which the various positions of the four consecutive proline-tyrosine motif and the five surrounding residues were replaced by all possible amino acid residues, we further elucidated the binding requirements of this motif.