Evidence for disulfide involvement in the regulation of intramolecular autolytic processing by human adamalysin19/ADAM19.
Evidence for disulfide involvement in the regulation of intramolecular autolytic processing by human adamalysin19/ADAM19.
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二硫键参与人 adamalysin19/ADAM19 分子内自溶过程调节的证据。
DOI：
10.1016/j.yexcr.2004.04.022
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发表时间：
2004
期刊：
影响因子：
--
通讯作者：
AmySang,Qing-Xiang
中科院分区：
文献类型：
--
作者：
Kang,Tiebang;Tschesche,Harald;AmySang,Qing-Xiang
人adamalysin 19（一种解整合素和金属蛋白酶19，hADAM19）通过弗林蛋白酶介导的前结构域裂解而被激活，随后在Glu586-Ser587富含半胱氨酸的结构域内进行自溶加工，该过程在分子内发生，产生与其COOH末端片段（C片段）相关的NH2末端片段（N片段），最有可能通过二硫键债券。当过表达可溶性 hADAM19 的稳定 Madin-Darby 犬肾 (MDCK) 转染子用二硫苏糖醇 (DTT) 或 pH 6.5、7.5 或 8.5 的培养基处理时，酶的分泌和折叠不受影响。自溶过程被 DTT 和 pH 6.5 介质阻断，有利于二硫键还原，但被 pH 8.5 介质增加，促进二硫键形成。 C 片段的 Cys605、Cys633、Cys639 和 Cys643 似乎部分负责 C 片段和 N 片段之间的共价缔合。当这些半胱氨酸残基单独突变为丝氨酸残基时，在可溶性突变体中鉴定出位于 Lys543–Val544 的新自溶加工位点。在稳定表达全长 Cys633Ser 突变体的 MDCK 细胞的培养基中也可检测到脱落片段。 Ilomastat/GM6001 抑制 hADAM19，IC50 为 447 nM，但几乎不影响脱落过程。富含半胱氨酸的结构域可能形成二硫键来调节 hADAM19 的自溶加工和脱落。
Human adamalysin 19 (a disintegrin and metalloproteinase 19, hADAM19) is activated by furin-mediated cleavage of the prodomain followed by an autolytic processing within the cysteine-rich domain at Glu586–Ser587, which occurs intramolecularly, producing an NH2terminal fragment (N-fragment) associated with its COOH-terminal fragment (C-fragment), most likely through disulfide bonds. When stable Madin-Darby canine kidney (MDCK) transfectants overexpressing soluble hADAM19 were treated with dithiothreitol (DTT) or with media at pH 6.5, 7.5, or 8.5, the secretion and folding of the enzyme were not affected. Autolytic processing was blocked by DTT and pH 6.5 media, which favor disulfide reduction, but was increased by pH 8.5 media, which promotes disulfide formation. Cys605, Cys633, Cys639, and Cys643of the C-fragment appear to be partially responsible for the covalent association between the C-fragment and the N-fragment. A new autolytic processing site at Lys543–Val544was identified in soluble mutants when these cysteine residues were individually mutated to serine residues. Shed fragments were also detectable in the media from MDCK cells stably expressing the full-length Cys633Ser mutant. Ilomastat/GM6001 inhibited hADAM19 with an IC50of 447 nM, but scarcely affected the shedding process. The cysteine-rich domain likely forms disulfide bonds to regulate the autolytic processing and shedding of hADAM19.
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影响因子：
3.1
作者：
Yun-Ge Zhao;Ping Wei;Q. Sang
通讯作者：
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影响因子：
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DOI：
--
发表时间：
2003
期刊：
--
影响因子：
--
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通讯作者：
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作者：
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影响因子：
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作者：
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