High-resolution analysis and functional mapping of cleavage sites and substrate proteins of furin in the human proteome.
High-resolution analysis and functional mapping of cleavage sites and substrate proteins of furin in the human proteome.
复制标题
DOI：
10.1371/journal.pone.0054290
复制
发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Cieplak P
中科院分区：
文献类型：
--
作者：
Shiryaev SA;Chernov AV;Golubkov VS;Thomsen ER;Chudin E;Chee MS;Kozlov IA;Strongin AY;Cieplak P
人们越来越认识到蛋白质水解过程在人类健康和疾病中的作用，但在蛋白质组范围内分析这些过程的工具有限。Furin是一种普遍存在的蛋白转化酶，它在基本残基后进行裂解，并将分泌的蛋白转化为具有生物活性的蛋白。尽管有这一重要作用，但在人类蛋白质组中仍有许多furin底物未知。我们设计了一种方法，用于蛋白酶目标鉴定，结合了一个在硅发现管道与高度多重的蛋白酶活性测定。我们对人类蛋白质组进行了计算机分析，确定了超过1050种分泌蛋白作为潜在的furin底物。然后，我们使用多重蛋白酶试验来验证这些暂定目标。该实验对超过3260个重叠肽进行了分析，这些重叠肽被设计为代表候选蛋白中furin切割位点的P7-P1 ‘和P4-P4 ’位置。获得的结果大大增加了我们对furin独特的裂解偏好的认识，揭示了短程（P4-P1）和远程（P7-P6）相互作用在定义furin裂解特异性中的重要性，证明了单独的R-X-R/K/X-R↓基序不足以预测furin蛋白水解的底物，并在人类蛋白质组中鉴定了约490种furin潜在的蛋白底物。这些底物在细胞通路上的分配表明furin在发育中起重要作用，包括轴突引导、心脏发生和干细胞多能性的维持。本研究提出的新方法可以很容易地应用于其他蛋白酶。
There is a growing appreciation of the role of proteolytic processes in human health and disease, but tools for analysis of such processes on a proteome-wide scale are limited. Furin is a ubiquitous proprotein convertase that cleaves after basic residues and transforms secretory proproteins into biologically active proteins. Despite this important role, many furin substrates remain unknown in the human proteome. We devised an approach for proteinase target identification that combines an in silico discovery pipeline with highly multiplexed proteinase activity assays. We performed in silico analysis of the human proteome and identified over 1,050 secretory proteins as potential furin substrates. We then used a multiplexed protease assay to validate these tentative targets. The assay was carried out on over 3,260 overlapping peptides designed to represent P7-P1’ and P4-P4’ positions of furin cleavage sites in the candidate proteins. The obtained results greatly increased our knowledge of the unique cleavage preferences of furin, revealed the importance of both short-range (P4-P1) and long-range (P7-P6) interactions in defining furin cleavage specificity, demonstrated that the R-X-R/K/X-R↓ motif alone is insufficient for predicting furin proteolysis of the substrate, and identified ∼490 potential protein substrates of furin in the human proteome. The assignment of these substrates to cellular pathways suggests an important role of furin in development, including axonal guidance, cardiogenesis, and maintenance of stem cell pluripotency. The novel approach proposed in this study can be readily applied to other proteinases.
登录
查看更多内容
影响因子：
3.7
作者：
Gordon WR;Vardar-Ulu D;L'Heureux S;Ashworth T;Malecki MJ;Sanchez-Irizarry C;McArthur DG;Histen G;Mitchell JL;Aster JC;Blacklow SC
通讯作者：
Blacklow SC
影响因子：
4.5
作者：
Deryugina, EI;Ratnikov, BI;Strongin, AY
通讯作者：
Strongin, AY
影响因子：
14.9
作者：
Jaroszewski L;Li Z;Cai XH;Weber C;Godzik A
通讯作者：
Godzik A
影响因子：
14.9
作者：
通讯作者：
--
DOI：
10.1111/j.2517-6161.1995.tb02031.x
发表时间：
1995-01-01
影响因子：
5.8
作者：
BENJAMINI, Y;HOCHBERG, Y
通讯作者：
HOCHBERG, Y