'Shed' furin: mapping of the cleavage determinants and identification of its C-terminus
'Shed' furin: mapping of the cleavage determinants and identification of its C-terminus
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DOI：
10.1042/0264-6021:3540689
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发表时间：
2001-03-15
影响因子：
4.1
通讯作者：
Schlokat, U
中科院分区：
文献类型：
--
作者：
Plaimauer, B;Mohr, G;Schlokat, U
人内切蛋白酶弗林蛋白酶参与多种生物活性蛋白质的前体分子的蛋白水解成熟。尽管其通过跨膜结构域定位在反式高尔基体系统的膜中，但已多次报道其形成C末端截短的天然分泌形式，称为“脱落”弗林蛋白酶。为了鉴定切割位点，引入了大小增加的内部缺失突变体，N-末端至Leu（708），随后引入了单个氨基酸取代，并鉴定Arg（683）为脱落的主要决定因素。MS分析确定Ser（682）为脱落弗林蛋白酶的C-末端，表明单碱基切割可能发生在Arg（683）的N-末端。Arg（683）的改变将脱落机制引导到先前未使用的替代切割位点。
The human endoprotease furin is involved in the proteolytic maturation of the precursor molecules of a wide variety of bioactive proteins. Despite its localization in the membranes of the trans-Golgi system by means of a transmembrane domain, it has repeatedly been reported to form a C-terminally truncated, naturally secreted form referred to as 'shed' furin. In order to identify the cleavage site, internal deletion mutants of increasing size, N-terminal to Leu(708), and subsequently individual amino acid substitutions were introduced, and Arg(683) was identified as the prime determinant for shedding. MS analysis determined Ser(682) as the C-terminus of shed furin, suggesting that monobasic cleavage may occur N-terminal to Arg(683). Alteration of Arg(683) directs the shedding mechanism to alternative cleaving sites previously unused.