A study of human furin specificity using synthetic peptides derived from natural substrates, and effects of potassium ions.
A study of human furin specificity using synthetic peptides derived from natural substrates, and effects of potassium ions.
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DOI：
10.1016/j.abb.2009.05.013
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发表时间：
2009-07-15
影响因子：
3.9
通讯作者：
Juliano L
中科院分区：
文献类型：
--
作者：
Izidoro MA;Gouvea IE;Santos JA;Assis DM;Oliveira V;Judice WA;Juliano MA;Lindberg I;Juliano L
我们探索弗林蛋白酶底物的要求，除了基序R-X-K/R-R使用合成的荧光共振能量转移（FRET）十肽。这些十肽衍生自病毒外壳糖蛋白以及人和细菌蛋白质前体中的弗林蛋白酶切割位点。大多数底物的弗林蛋白酶的水解被激活的K+离子，而亲液性的Hofmeister系列的阴离子的抑制剂。弗林蛋白酶水解活性的分析表明，其效率是高度依赖于特定的组合的氨基酸在不同的底物位置。弗林蛋白酶亚位点之间存在明显的相互依赖性，在确定其特异性和设计抑制剂时必须考虑到这一点。然而，在P′1处检测到对S的明显偏好，在P′2处检测到对V的明显偏好，在P′3处，氨基酸D、S、L和A的频率几乎相等。在非主要亚位点中，最佳底物在P6呈现S和H;在P5呈现碱性氨基酸;在P3没有明显趋势。有趣的是，在源自H5 N1流感血凝素弗林蛋白酶加工位点的肽的主要侧上的两个氨基酸取代大大改善了其水解。这些修改是可能的单点突变，这表明一个更具感染性的病毒的潜在产量。
We explored furin substrate requirements in addition to the motif R-X-K/R-R using synthetic fluorescent resonance energy transfer (FRET) decapeptides. These decapeptides were derived from furin cleavage sites in viral coat glycoproteins and human and bacterial protein precursors. The hydrolysis by furin of most substrate was activated by K+ ion, whereas kosmotropic anions of the Hofmeister series were inhibitors. The analysis of furin hydrolytic activity showed that its efficiency is highly dependent on the particular combinations of amino acids at different substrate positions. There is a clear interdependence of furin subsites that must be taken in account in determining its specificity and also for the design of inhibitors. However, clear preferences were detected for substrates with S at P′1, and V at P′2, at P′3 the amino acids D, S, L and A are almost equally frequent. In the non-prime subsites the best substrates presented S and H at P6; basic amino acids at P5; and no clear tendency at P3. Interestingly, two amino acid substitutions on the prime side of the peptide derived from H5N1 influenza hemagglutinin furin processing site highly improved its hydrolysis. These modifications are possible by single point mutations, suggesting a potential yield of a more infectious virus.
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DOI：
10.1038/nsb941
发表时间：
2003-07-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
--
作者：
Henrich, S;Cameron, A;Than, ME
通讯作者：
Than, ME
影响因子：
3.5
作者：
GU, ML;RAPPAPORT, J;LEPPLA, SH
通讯作者：
LEPPLA, SH
影响因子：
2.9
作者：
ANGLIKER, H;NEUMANN, U;THOMAS, G
通讯作者：
THOMAS, G
影响因子：
5.4
作者：
Binley, JM;Sanders, RW;Moore, JP
通讯作者：
Moore, JP
影响因子：
4.8
作者：
Krysan, DJ;Rockwell, NC;Fuller, RS
通讯作者：
Fuller, RS