Characterization of an aldolase-binding site in the Wiskott-Aldrich syndrome protein
Characterization of an aldolase-binding site in the Wiskott-Aldrich syndrome protein
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DOI：
10.1074/jbc.m506346200
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发表时间：
2006-01-20
影响因子：
4.8
通讯作者：
Nussenzweig, V
中科院分区：
文献类型：
--
作者：
Buscaglia, CA;Penesetti, D;Nussenzweig, V
血栓反应蛋白相关匿名蛋白（TRAP）是疟原虫孢子体中必不可少的跨膜分子。TRAP在细胞外部分显示粘附基元，而其细胞质尾部通过醛缩酶与肌动蛋白连接，从而驱动寄生虫运动和宿主细胞入侵。对TRAP与醛缩酶结合的最低要求进行了扫描，发现不同的人类蛋白质共有，包括Wiskott-Aldrich综合征蛋白（WASp）家族成员。在体外和体内，通过生化、缺失定位、诱变和共免疫沉淀研究来表征WASp成员与醛缩酶的结合。与TRAP的情况一样，WASp与醛缩酶的结合被酶底物/产物竞争性地抑制。此外，TRAP和WASp，而不是其他不相关的醛缩酶结合物，在体外竞争与酶的结合。总之，我们的研究结果在WASp家族成员中定义了一个保守的醛缩酶结合基序，并表明醛缩酶调节哺乳动物细胞的运动性和肌动蛋白动力学。这些发现以及在其他人类蛋白质中存在类似的醛缩酶结合基序，其中一些在拉下实验中确实与醛缩酶相互作用，表明该酶具有补充的非糖酵解作用。
The thrombospondin-related anonymous protein (TRAP) is an essential transmembrane molecule in Plasmodium sporozoites. TRAP displays adhesive motifs on the extracellular portion, whereas its cytoplasmic tail connects to actin via aldolase, thus driving parasite motility and host cell invasion. The minimal requirements for the TRAP binding to aldolase were scanned here and found to be shared by different human proteins, including the Wiskott-Aldrich syndrome protein (WASp) family members. In vitro and in vivo binding of WASp members to aldolase was characterized by biochemical, deletion mapping, mutagenesis, and co-immunoprecipitation studies. As in the case of TRAP, the binding of WASp to aldolase is competitively inhibited by the enzyme substrate/products. Furthermore, TRAP and WASp, but not other unrelated aldolase binders, compete for the binding to the enzyme in vitro. Together, our results define a conserved aldolase binding motif in the WASp family members and suggest that aldolase modulates the motility and actin dynamics of mammalian cells. These findings along with the presence of similar aldolase binding motifs in additional human proteins, some of which indeed interact with aldolase in pull-down assays, suggest supplementary, non-glycolytic roles for this enzyme.