Development of a Novel Tetravalent Synthetic Peptide That Binds to Phosphatidic Acid.
Development of a Novel Tetravalent Synthetic Peptide That Binds to Phosphatidic Acid.
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DOI：
10.1371/journal.pone.0131668
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Umeda M
中科院分区：
文献类型：
--
作者：
Ogawa R;Nagao K;Taniuchi K;Tsuchiya M;Kato U;Hara Y;Inaba T;Kobayashi T;Sasaki Y;Akiyoshi K;Watanabe-Takahashi M;Nishikawa K;Umeda M
我们采用多价肽库筛选技术来鉴定与磷脂酸（PA）结合的肽基序，而不是与磷脂酰胆碱（PC），磷脂酰乙醇胺（PE）和磷脂酰丝氨酸（PS）结合的肽基序。一种具有MARWHRHHH序列基序的四价肽，被称为pabp - tp（磷脂酸结合四价肽），在由PC和胆固醇组成的双层膜中可以结合低至1mol %的PA。对苯胺- tp与含10 mol%苯胺的膜相互作用的动力学分析表明，苯胺- tp与苯胺的解离常数较低，KD = 38±5 nM。膜中胆固醇或PE与PA共存，通过增加磷酸单酯头基团的电离以及改变膜中PA分子的微环境，增强了PA - tp与PA的结合。氨基酸替代分析表明，PA - tp的第4位色氨酸残基参与了与PA的相互作用。此外，在PA - tp的第5至9位的一系列氨基酸替换表明，连续的组氨酸和精氨酸残基参与了PA磷酸单酯头基的识别。我们的研究结果表明，PAB-TP对PA的识别是通过疏水、静电和氢键相互作用的结合来实现的，并且PAB-TP的四价结构有助于在膜中与PA的高亲和力结合。新的PA结合四价肽PA- tp将深入了解PA结合蛋白识别PA的分子机制，PA结合蛋白参与各种细胞事件。
We employed a multivalent peptide-library screening technique to identify a peptide motif that binds to phosphatidic acid (PA), but not to other phospholipids such as phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylserine (PS). A tetravalent peptide with the sequence motif of MARWHRHHH, designated as PAB-TP (phosphatidic acid-binding tetravalent peptide), was shown to bind as low as 1 mol% of PA in the bilayer membrane composed of PC and cholesterol. Kinetic analysis of the interaction between PAB-TP and the membranes containing 10 mol% of PA showed that PAB-TP associated with PA with a low dissociation constant of KD = 38 ± 5 nM. Coexistence of cholesterol or PE with PA in the membrane enhanced the PAB-TP binding to PA by increasing the ionization of the phosphomonoester head group as well as by changing the microenvironment of PA molecules in the membrane. Amino acid replacement analysis demonstrated that the tryptophan residue at position 4 of PAB-TP was involved in the interaction with PA. Furthermore, a series of amino acid substitutions at positions 5 to 9 of PAB-TP revealed the involvement of consecutive histidine and arginine residues in recognition of the phosphomonoester head group of PA. Our results demonstrate that the recognition of PA by PAB-TP is achieved by a combination of hydrophobic, electrostatic and hydrogen-bond interactions, and that the tetravalent structure of PAB-TP contributes to the high affinity binding to PA in the membrane. The novel PA-binding tetravalent peptide PAB-TP will provide insight into the molecular mechanism underlying the recognition of PA by PA-binding proteins that are involved in various cellular events.
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DOI：
10.1016/j.bbalip.2013.10.005
发表时间：
2014-01
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
Nagao K;Hata M;Tanaka K;Takechi Y;Nguyen D;Dhanasekaran P;Lund-Katz S;Phillips MC;Saito H
通讯作者：
Saito H
DOI：
10.1073/pnas.93.23.12867
发表时间：
1996-11-12
影响因子：
11.1
作者：
Emoto, K;Kobayashi, T;Umeda, M
通讯作者：
Umeda, M
影响因子：
3.3
作者：
Nakanishi, H;de los Santos, P;Neiman, AM
通讯作者：
Neiman, AM
影响因子：
56.9
作者：
Loewen, CJR;Gaspar, ML;Levine, TP
通讯作者：
Levine, TP
影响因子：
4.8
作者：
Ghosh, S;Moore, S;Dush, M
通讯作者：
Dush, M