Redefining the specificity of phosphoinositide-binding by human PH domain-containing proteins.
Redefining the specificity of phosphoinositide-binding by human PH domain-containing proteins.
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DOI：
10.1038/s41467-021-24639-y
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发表时间：
2021-07-15
影响因子：
16.6
通讯作者：
Chen J
中科院分区：
文献类型：
--
作者：
Singh N;Reyes-Ordoñez A;Compagnone MA;Moreno JF;Leslie BJ;Ha T;Chen J
Pleckstrin同源结构域（PH）被认为与磷酸肌苷（PIPs）结合，但大多数含PH结构域的蛋白质与PIPs的特异性相互作用和PIPs的调节尚不清楚。在这里，我们采用单分子下拉实验来研究哺乳动物全细胞裂解物中脂质囊泡与全长蛋白的相互作用。在最初检测的67种人类PH结构域蛋白中，发现36种（54%）与PIPs具有不同特异性的亲和力，其中大多数以前未被报道过。对ARHGEF3的进一步研究揭示了其与PI(4,5)P2和PI(3,5)P2结合的不同结构要求，以及其与PI(4,5)P2结合的功能相关性。我们基于检测结果生成递归学习算法来分析242个人类PH结构域的序列，预测其中49%与pip结合。对20种预测的结合物和11种预测的非结合物进行了分析，结果与预测高度一致。综上所述，我们的发现揭示了PH结构域蛋白意想不到的脂质结合特异性。Pleckstrin同源结构域（PH）被认为与磷酸肌苷（pip）结合，但只有少数特定的相互作用是已知的。通过基于TIRF显微镜的分析、功能研究和实验验证的预测算法，作者表明特异性PIP结合在人类PH结构域中广泛存在。
Pleckstrin homology (PH) domains are presumed to bind phosphoinositides (PIPs), but specific interaction with and regulation by PIPs for most PH domain-containing proteins are unclear. Here we employ a single-molecule pulldown assay to study interactions of lipid vesicles with full-length proteins in mammalian whole cell lysates. Of 67 human PH domain-containing proteins initially examined, 36 (54%) are found to have affinity for PIPs with various specificity, the majority of which have not been reported before. Further investigation of ARHGEF3 reveals distinct structural requirements for its binding to PI(4,5)P2 and PI(3,5)P2, and functional relevance of its PI(4,5)P2 binding. We generate a recursive-learning algorithm based on the assay results to analyze the sequences of 242 human PH domains, predicting that 49% of them bind PIPs. Twenty predicted binders and 11 predicted non-binders are assayed, yielding results highly consistent with the prediction. Taken together, our findings reveal unexpected lipid-binding specificity of PH domain-containing proteins. Pleckstrin homology (PH) domains are presumed to bind phosphoinositides (PIPs) but only few specific interactions are known. Using TIRF microscopy based assays, functional studies and an experimentally validated prediction algorithm, the authors show that specific PIP binding is widespread among human PH domains.
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影响因子：
16
作者：
Park, Wei Sun;Do Heo, Won;Teruel, Mary N.
通讯作者：
Teruel, Mary N.
影响因子：
14.9
作者：
Mistry J;Chuguransky S;Williams L;Qureshi M;Salazar GA;Sonnhammer ELL;Tosatto SCE;Paladin L;Raj S;Richardson LJ;Finn RD;Bateman A
通讯作者：
Bateman A
DOI：
10.1016/j.bbalip.2015.02.013
发表时间：
2015-06
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
Hammond GR;Balla T
通讯作者：
Balla T
影响因子：
21.3
作者：
Kanai, F;Liu, H;Yaffe, MB
通讯作者：
Yaffe, MB
DOI：
10.1073/pnas.92.23.10472
发表时间：
1995-11-07
影响因子：
11.1
作者：
LEMMON, MA;FERGUSON, KM;SCHLESSINGER, J
通讯作者：
SCHLESSINGER, J