Characterization of the tetratricopeptide-containing domain of BUB1, BUBR1, and PP5 proves that domain amphiphilicity over amino acid sequence specificity governs protein adsorption and interfacial activity
Characterization of the tetratricopeptide-containing domain of BUB1, BUBR1, and PP5 proves that domain amphiphilicity over amino acid sequence specificity governs protein adsorption and interfacial activity
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DOI：
10.1021/jp711222s
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发表时间：
2008-07-10
影响因子：
3.3
通讯作者：
Bolanos-Garcia, Victor M.
中科院分区：
文献类型：
--
作者：
Beaufils, Sylvie;Grossmann, J. Guenter;Bolanos-Garcia, Victor M.
四肽基序重复（TPR）是通常介导蛋白质-蛋白质并且在某些情况下介导蛋白质-脂质相互作用的α-螺旋-转角-α-螺旋基序。由于它的成功，这个基序在进化过程中被保留下来，并且可以在低等和高等生物的广泛功能的蛋白质中被识别。BUB 1、BUBR 1和蛋白磷酸酶5（PP 5）的N-末端区域含有TPR基序的串联排列。BUB 1和BUBR 1是保守的多结构域蛋白激酶，在有丝分裂检查点中发挥关键作用，该机制确保染色体分离的同步性。PP 5是一种靶向广泛蛋白质底物的酶，包括单跨膜受体和哺乳动物隐花色素。PP 5的N-末端TPR结构域通过与蛋白质和脂质分子的直接相互作用来调节C-末端催化结构域的活性。我们描绘的TPR基序的BUB 1，BUBR 1，和PP 5的串联安排的生物物理和生物化学性质，使用远紫外光谱，解决方案X-射线散射，零椭圆偏振，表面流变学测量，和布鲁斯特角显微镜（BAM）的意见。我们发现，尽管低的氨基酸序列保守性和不同的功能，TPR基序重复的三种蛋白质表现出类似的界面特性，包括吸附动力学，高表面活性，并在空气/水界面形成稳定的，刚性的薄膜。我们的研究表明，结构域的两亲性是更重要的比氨基酸序列的特异性，在确定蛋白质吸附和界面活性。
The tetratricopeptide motif repeat (TPR) is an alpha-helix-turn-alpha-helix motif that typically mediates protein -protein and, in some cases, protein-lipid interactions. Because of its success, this motif has been preserved through evolution and can be identified in proteins of a wide range of functions in lower and higher organisms. The N-terminal region of BUB1, BUBR1, and protein phosphatase 5 (PP5) contains tandem arrangements of the TPR motif. BUB1 and BUBR1 are conserved multidomain protein kinases that play a key role in the mitotic checkpoint, the mechanism that ensures the synchrony of chromosome segregation. PP5 is an enzyme that targets a wide range of protein substrates including single transmembrane receptors and mammalian cryptochromes. The N-terminal TPR domain of PP5 regulates the activity of the C-terminal catalytic domain through direct interaction with protein and lipid molecules. We portray the biophysical and biochemical properties of the tandem arrangements of the TPR motif of BUB1, BUBR1, and PP5 using far-UV spectroscopy, solution X-ray scattering, null ellipsometry, surface theology measurements, and Brewster angle microscopy (BAM) observations. We show that, despite the low amino acid sequence conservation and different function, the TPR motif repeats of the three proteins exhibit similar interfacial properties including adsorption kinetics, high surface activity, and the formation of stable, rigid films at the air/water interface. Our studies demonstrate that domain amphiphilicity is of higher importance than amino acid sequence specificity in the determination of protein adsorption and interfacial activity.