Calprotectin inhibits matrix metalloproteinases by sequestration of zinc
Calprotectin inhibits matrix metalloproteinases by sequestration of zinc
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DOI：
10.1136/mp.54.5.289
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发表时间：
2001-10-01
期刊：
影响因子：
--
通讯作者：
Fagerhol, MK
中科院分区：
文献类型：
--
作者：
Isaksen, B;Fagerhol, MK
背景/目的钙保护素是一种存在于中性粒细胞胞浆中的36 kDa蛋白质，具有抗菌和诱导细胞凋亡的活性，而锌的加入则逆转了这一活性。基质金属蛋白酶(MMPs)是一类锌依赖酶家族，在胚胎发育、血管生成、伤口愈合等许多正常的生物学过程中起重要作用，也在炎症、癌症和组织破坏等病理过程中发挥重要作用。本研究的目的是研究钙保护素是否能抑制基质金属蛋白酶的活性，以及这种抑制作用是否可以被锌的加入所克服。方法-基质金属蛋白酶的活性通过预涂在微孔上的底物的降解来测量，并通过剩余底物的考马斯亮蓝染色来显示。以明胶和α-酪蛋白为底物，检测了7种金属蛋白酶(MMP1、MMP2、MMP3、MMP7、MMP8、MMP9和MMP13)对明胶和α-酪蛋白的抑制活性。钙保护素的加入抑制了所有MMPs的活性，但从0.3um到~gt；11um的不同浓度的蛋白质对MMPs的抑制率为50%。钙保护素的抑制作用在很大程度上被锌的加入所克服。结论--研究结果表明，钙保护素通过隔离锌来抑制MMPs。数据还表明，MMPs与锌的亲和力不同，而钙保护素对锌的亲和力低于MMPs。
Background/Aimss-Calprotectin, a 36 kDa protein present in neutrophil cytoplasm, has antimicrobial and apoptosis inducing activities, which are reversed by the addition of zinc. Matrix metalloproteinases (MMPs), a family of zinc dependent enzymes, are important in many normal biological processes including embryonic development, angiogenesis, and wound healing, but also pathological processes such as inflammation, cancer, and tissue destruction. The aim of this study was to investigate whether calprotectin can inhibit MMP activity, and whether such inhibition could be overcome by the addition of zinc.Methods-MMP activity was measured by the degradation of substrates precoated on to microwells, and visualised by Coomassie blue staining of residual substrate. Seven metalloproteinases (MMP-1, MMP-2, MMP-3, MMP-7, MMP-8, MMP-9, and MMP-13) were teste against two substrates: gelatin and alpha -casein.Results-All MMPs except MMP-1 were active against gelatin, whereas MMP-7 was the only enzyme active against alpha -casein. The addition of calprotectin inhibited the activity of all the MMPs, but different concentrations of the protein, from 0.3 muM to > 11 muM, were necessary to produce a 50% inhibition of the MMPs. Inhibition by calprotectin was largely overcome by the addition of zinc.Conclusions-The findings suggest that calprotectin inhibits MMPs by sequestration of zinc. The data also suggest that MMPs have different affinities for zinc and that calprotectin has a lower zinc affinity than the MMPs.