The high molecular weight urinary matrix metalloproteinase (MMP) activity is a complex of gelatinase B/MMP-9 and neutrophil gelatinase-associated lipocalin (NGAL) - Modulation of MMP-9 activity by NGAL
The high molecular weight urinary matrix metalloproteinase (MMP) activity is a complex of gelatinase B/MMP-9 and neutrophil gelatinase-associated lipocalin (NGAL) - Modulation of MMP-9 activity by NGAL
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DOI：
10.1074/jbc.m106089200
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发表时间：
2001-10-05
影响因子：
4.8
通讯作者：
Moses, MA
中科院分区：
文献类型：
--
作者：
Yan, L;Borregaard, N;Moses, MA
多种癌症患者尿液中基质金属蛋白酶（MMP）活性的检测与疾病状态密切相关。在这些活性中，一组高分子量（HAM） MMPs的存在独立地作为转移表型的多变量预测因子(1)。尽管这些HMW - MMP活性具有新颖性，并且在非侵袭性癌症诊断和/或预后方面具有潜在的重要应用，但它们的身份仍然未知。在这里，我们报道了一种类似125-kDa的HMW尿MMPs的鉴定，它是明胶酶B （MMP-9）和中性粒细胞明胶酶相关脂钙蛋白（NGAL）的复合物。多种生物化学方法证实了这一身份。底物凝胶电泳分析表明，125 kda的尿MMP活性与纯化的人中性粒细胞MMP-9共迁移。NGAL复杂。125kda尿MMP9。NGAL复合物被纯化的抗人NGAL抗体和单特异性抗人MMP-9抗体识别。此外，这两种抗体能够以剂量依赖的方式独立地特异性免疫沉淀125 kda尿MMP活性。此外，MMP-9的复合物。将MMP-9和NGAL混合在pH值在尿和正常尿范围内的明胶酶缓冲液中，可以在体外重建NGAL。最后，利用重组人NGAL和MMP-9在体外以及在人乳腺癌细胞中过表达NGAL进行细胞培养，研究了NGAL和MMP-9相互作用的生化后果。我们的数据表明，NGAL能够以剂量依赖的方式保护MMP-9免受降解，从而保持MMP-9的酶活性。总之，本研究确定了125 kda尿明胶酶是MMP-9和NGAL的复合物，并提供了NGAL通过保护MMP-9免受降解而调节其活性的证据。
Detection of matrix metalloproteinase (MMP) activities in the urine from patients with a variety of cancers has been closely correlated to disease status. Among these activities, the presence of a group of high molecular weight (HAM) MMPs independently serves as a multivariate predictor of the metastatic phenotype (1). The identity of these HMW MMP activities has remained unknown despite their novelty and their potentially important applications in non-invasive cancer diagnosis and/or prognosis. Here, we report the identification of one of these HMW urinary MMPs of similar to 125-kDa as being a complex of gelatinase B (MMP-9) and neutrophil gelatinase-associated lipocalin (NGAL). Multiple biochemical approaches verified this identity. Analysis using substrate gel electrophoresis demonstrated that the 125-kDa urinary MMP activity co-migrates with purified human neutrophil MMP-9.NGAL complex. The 125-kDa urinary MMP9.NGAL complex was recognized by a purified antibody against human NGAL as well as by a monospecific antihuman MMP-9 antibody. Furthermore, these same two antibodies were independently capable of specifically immunoprecipitating the 125-kDa urinary MMP activity in a dose-dependent manner. In addition, the complex of MMP-9.NGAL could be reconstituted in vitro by mixing MMP-9 and NGAL in gelatinase buffers with pH values in the range of urine and in normal urine as well. Finally, the biochemical consequences of the NGAL and MMP-9 interaction were investigated both in vitro using recombinant human NGAL and MMP-9 and in cell culture by overexpressing NGAL in human breast carcinoma cells. Our data demonstrate that NGAL is capable of protecting MMP-9 from degradation in a dose-dependent manner and thereby preserving MMP-9 enzymatic activity. In summary, this study identifies the 125-kDa urinary gelatinase as being a complex of MMP-9 and NGAL and provides evidence that NGAL modulates MMP-9 activity by protecting it from degradation.