Membrane Interactions of the Peroxisomal Proteins PEX5 and PEX14.
Membrane Interactions of the Peroxisomal Proteins PEX5 and PEX14.
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DOI：
10.3389/fcell.2021.651449
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发表时间：
2021
影响因子：
5.5
通讯作者：
Sattler M
中科院分区：
文献类型：
--
作者：
Gaussmann S;Gopalswamy M;Eberhardt M;Reuter M;Zou P;Schliebs W;Erdmann R;Sattler M
人PEX 5和PEX 14是过氧化物酶体转位子的基本组分，其介导货物酶输入过氧化物酶体。PEX 5是货物酶的可溶性受体，由N-末端固有无序结构域（NTD）和C-末端三肽（TPR）结构域组成，其识别货物蛋白中的过氧化物酶体靶向信号1（PTS 1）肽基序。PEX 5 NTD含有多个WF肽基序（WxxxF/Y或相关基序），这些基序被膜相关蛋白PEX 14的NTD中的小球状结构域识别。PEX 5或PEX 14 NTD如何与过氧化物酶体膜结合以及两种蛋白质之间的相互作用如何在膜上调节尚不清楚。在这里，我们的PEX 5 NTD和PEX 14 NTD在体外的膜相互作用的特点，通过膜模仿bicelles和nanodisces使用NMR光谱和等温滴定量热法。PEX 14 NTD与具有与WxxxF/Y结合位点部分重叠的表面的膜模拟双胞弱相互作用。PEX 5 NTD具有多个与膜相互作用的位点，涉及许多两亲性α-螺旋区域，其中包括一些WxxxF/Y基序。这些区域的部分形成的α-螺旋构象在bicelles存在下稳定。值得注意的是，ITC数据显示，PEX 5和PEX 14 NTD之间的相互作用在很大程度上不受膜存在的影响。PEX 5/PEX 14相互作用表现出类似的自由结合焓，其中在双胞存在下降低的结合焓由降低的熵损失补偿。这表明PEX 5与PEX 14在膜上的对接不会降低两种蛋白质之间的总体结合亲和力，从而提供了对过氧化物酶体易位子组装中PEX 5-PEX 14对接的初始阶段的了解。
Human PEX5 and PEX14 are essential components of the peroxisomal translocon, which mediates import of cargo enzymes into peroxisomes. PEX5 is a soluble receptor for cargo enzymes comprised of an N-terminal intrinsically disordered domain (NTD) and a C-terminal tetratricopeptide (TPR) domain, which recognizes peroxisomal targeting signal 1 (PTS1) peptide motif in cargo proteins. The PEX5 NTD harbors multiple WF peptide motifs (WxxxF/Y or related motifs) that are recognized by a small globular domain in the NTD of the membrane-associated protein PEX14. How the PEX5 or PEX14 NTDs bind to the peroxisomal membrane and how the interaction between the two proteins is modulated at the membrane is unknown. Here, we characterize the membrane interactions of the PEX5 NTD and PEX14 NTD in vitro by membrane mimicking bicelles and nanodiscs using NMR spectroscopy and isothermal titration calorimetry. The PEX14 NTD weakly interacts with membrane mimicking bicelles with a surface that partially overlaps with the WxxxF/Y binding site. The PEX5 NTD harbors multiple interaction sites with the membrane that involve a number of amphipathic α-helical regions, which include some of the WxxxF/Y-motifs. The partially formed α-helical conformation of these regions is stabilized in the presence of bicelles. Notably, ITC data show that the interaction between the PEX5 and PEX14 NTDs is largely unaffected by the presence of the membrane. The PEX5/PEX14 interaction exhibits similar free binding enthalpies, where reduced binding enthalpy in the presence of bicelles is compensated by a reduced entropy loss. This demonstrates that docking of PEX5 to PEX14 at the membrane does not reduce the overall binding affinity between the two proteins, providing insights into the initial phase of PEX5-PEX14 docking in the assembly of the peroxisome translocon.
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影响因子：
16.1
作者：
Chen, Xiaoying;Zaro, Jennica L.;Shen, Wei-Chiang
通讯作者：
Shen, Wei-Chiang
DOI：
10.1083/jcb.135.1.1
发表时间：
1996-10
期刊：
The Journal of cell biology
影响因子：
--
作者：
Distel B;Erdmann R;Gould SJ;Blobel G;Crane DI;Cregg JM;Dodt G;Fujiki Y;Goodman JM;Just WW;Kiel JA;Kunau WH;Lazarow PB;Mannaerts GP;Moser HW;Osumi T;Rachubinski RA;Roscher A;Subramani S;Tabak HF;Tsukamoto T;Valle D;van der Klei I;van Veldhoven PP;Veenhuis M
通讯作者：
Veenhuis M
影响因子：
14.8
作者：
Hagn F;Nasr ML;Wagner G
通讯作者：
Wagner G
影响因子：
3.9
作者：
PICK, U
通讯作者：
PICK, U
影响因子：
19
作者：
Erdmann, R;Veenhuis, M;Kunau, WH
通讯作者：
Kunau, WH