N-arginine dibasic convertase (nardilysin) isoforms are soluble dibasic-specific metalloendopeptidases that localize in the cytoplasm and at the cell surface.

N-arginine dibasic convertase (nardilysin) isoforms are soluble dibasic-specific metalloendopeptidases that localize in the cytoplasm and at the cell surface.
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N-精氨酸二碱基转化酶(nardilysin)同工型是可溶性二碱基特异性金属内肽酶,位于细胞质和细胞表面。

DOI:
10.1042/bj3490587
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发表时间:
2000
期刊:
The Biochemical journal
影响因子:
--
通讯作者:
A. Prat
A. Prat
中科院分区:
--
文献类型:
--
作者:
V. Hospital;V. Chesneau;Á. Balogh;C. Joulie;N. Seidah;P. Cohen;A. Prat

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N-精氨酸(R)二元(NRD)转化酶(nardilysin; EC 3.4.24.61),一种M16家族的金属内肽酶,在体外特异性切割二元基序中精氨酸N-末端的肽底物。在大鼠睾丸中,该酶定位于精子细胞的细胞质内,并与轴丝和轴柄的微管相关联。NRD 1和NRD 2转化酶是两种NRD转化酶亚型,其区别在于在活性位点附近不存在(亚型1)或存在(亚型2)68个氨基酸的插入。在这项研究中,我们过表达这两种亚型,无论是通过牛痘病毒感染的BSC 40细胞或转染COS-7细胞。部分纯化的酶表现出非常相似的生化和酶学性质。微测序显示NRD转化酶是N-末端加工的。免疫细胞荧光,免疫电镜和亚细胞分级分离研究的结果主张在一个主要的两种肽酶的胞质定位。虽然推定的信号肽没有直接NRD转化酶在体外翻译试验中进入微粒体,生物素化实验清楚地表明,在细胞表面的两种亚型的存在。总之,虽然大多数已知的加工事件在对碱性残基的分泌途径内的前蛋白转化酶实现,NRD转化酶可以履行类似的功能,在细胞质和/或在细胞表面。
N-arginine (R) dibasic (NRD) convertase (nardilysin; EC 3.4.24.61), a metalloendopeptidase of the M16 family, specifically cleaves peptide substrates at the N-terminus of arginines in dibasic motifs in vitro. In rat testis, the enzyme localizes within the cytoplasm of spermatids and associates with microtubules of the manchette and axoneme. NRD1 and NRD2 convertases, two NRD convertase isoforms, differ by the absence (isoform 1) or presence (isoform 2) of a 68-amino acid insertion close to the active site. In this study, we overexpressed both isoforms, either by vaccinia virus infection of BSC40 cells or transfection of COS-7 cells. The partially purified enzymes exhibit very similar biochemical and enzymic properties. Microsequencing revealed that NRD convertase is N-terminally processed. Results of immunocytofluorescence, immunoelectron microscopy and subcellular fractionation studies argue in favour of a primary cytosolic localization of both peptidases. Although the putative signal peptide did not direct NRD convertase into microsomes in an in vitro translation assay, biotinylation experiments clearly showed the presence of both isoforms at the cell surface. In conclusion, although most known processing events at pairs of basic residues are achieved by proprotein convertases within the secretory pathway, NRD convertase may fulfil a similar function in the cytoplasm and/or at the cell surface.
参与前体成熟的叶绿体加工酶与最近认识的金属内肽酶家族共享锌结合基序。
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