Characterization of the catalytic activity of the membrane-anchored metalloproteinase ADAM15 in cell-based assays

Maretzky, Thorsten; Yang, Guangli; Ouerfelli, Ouathek; Overall, Christopher M.; Worpenberg, Susanne; Hassiepen, Ulrich; Eder, Joerg; Blobel, Carl P.

doi:10.1042/bj20082127

Cited by 47 publications

(43 citation statements)

References 53 publications

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“…Fibroblasts and MCF-7 cells were transfected with Lipofectamine 2000, and COS7 cells with Lipofectamine, as described (6). Stable cell lines were selected in medium containing either 50 to 200 Ag/mL hygromycin B or 25 to 100 Ag/mL zeocin (Invitrogen).…”

Section: Methodsmentioning

confidence: 99%

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Src Stimulates Fibroblast Growth Factor Receptor-2 Shedding by an ADAM15 Splice Variant Linked to Breast Cancer

et al. 2009

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ADAMs (a disintegrin and metalloproteinase) have important roles in development and diseases such as cancer. Previously, an ADAM15 splice variant (ADAM15B), which contains an inserted cytoplasmic Src-binding site, was linked to clinical aggressiveness in breast cancer, yet little was known about how this splice variant affects the function of ADAM15. Here, we show that ADAM15B has enhanced catalytic activity in cellbased assays compared with ADAM15A, which lacks a Srcbinding site, using shedding of fibroblast growth factor receptor 2iiib variant as an assay for catalytic activity. Moreover, the enhanced activity of ADAM15B compared with ADAM15A depends on Src because it is abolished by Srckinase inhibitors and in Src À/À cells, but not in Src À/À cells rescued with Src. These findings provide insights into the mechanism of how a splice variant linked to clinical agressiveness in breast cancer causes increased activity of ADAM15B, and suggest that inhibitors of the ADAM15 protease activity or of the interaction of ADAM15B with Src could be useful to treat breast cancer in patients with dysregulated ADAM15B. [Cancer Res 2009;69(11):4573-6]

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Section: Methodsmentioning

confidence: 99%

“…Cells were transfected with FGFR2-AP and washed with OptiMEM 1 d after transfection; fresh OptiMEM with or without inhibitors of Src-kinases was added and incubated for 1 to 4 h, as indicated (6). AP activity in the supernatant and cell lysates of at least three identically treated wells was measured by colorimetry as described (6,11).…”

Section: Methodsmentioning

confidence: 99%

Src Stimulates Fibroblast Growth Factor Receptor-2 Shedding by an ADAM15 Splice Variant Linked to Breast Cancer

et al. 2009

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“…TIMP-3 is particularly effective at inhibiting MMP-1 and -2, and the membrane-bound MMPs [1,2]. Among the four TIMPs, TIMP-3 is the only one to additionally inhibit a large variety of disintegrin-metalloproteinases (ADAM, particularly ADAM-10, -12, -17, -28, and - 33) and disintegrin-metalloproteinases with ThromboSpondinlike motifs (ADAMTS, principally ADAMTS-1, -4, and -5), which help direct cell functions such as proliferation and migration [1,3,4]. Due to its role in regulating MMP, ADAM, and ADAMTS activity and its unique property to reside exclusively in the ECM, TIMP-3 may strongly influence hematopoiesis and ECM remodeling [5].…”

Section: Introductionmentioning

confidence: 99%

Altered Bone Mechanics, Architecture and Composition in the Skeleton of TIMP-3-Deficient Mice

et al. 2017

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Tissue inhibitor of metalloproteinases-3 (TIMP-3) maintains a healthy extracellular matrix by regulating matrix metalloproteinases (MMP), disintegrin-metalloproteinases (ADAM), and disintegrin-metalloproteinases with ThromboSpondin-like motifs (ADAMTS) activity. Currently, there is a need for a comprehensive understanding of the effects of TIMP-3 on the bone quality and integrity. In this study, we examined the mechanical, morphological, and compositional properties of TIMP-3 knock out (Timp-3 ) mouse bone. We hypothesize that the lack of TIMP-3 plays an important role in maintaining the overall bone integrity. Mechanical properties of humeri, lumbar vertebrae, and femurs from Timp-3 mice were determined using 3-point bending, compression, and notched 3-point bending, respectively. Morphological properties of the humeral cortical and trabecular bone and the caudal vertebrae cortical bone were evaluated using micro-computed tomography, while the composition of the femoral cortical and trabecular bone was examined using Fourier transform infrared spectroscopic imaging. Our results revealed that the integrity of the Timp-3 bone is compromised due to changes in its composition, structure, and mechanics. Reductions in the yield and ultimate load and stress capacity, and loss in bone fracture toughness were attributed to reduced density and thickness, and increased porosity of cortical bone. Thin trabeculae were dense, highly connected, and closely packed in Timp-3 bone. Furthermore, altered cortical and trabecular bone mineralization and increased compositional heterogeneity were found in Timp-3 bone, all being indicative of high bone remodeling. In conclusion, this study suggests that the lack of TIMP-3 is detrimental to bone development and maintenance.

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“…ADAM15 plays a role in endothelial barrier function, which suggests a possible role in lung inflammation. As a protease, ADAM15 can remove fibroblast growth factor receptor II components from the ECM and possibly also E-cadherin (111,113,126). ADAM15 also contains an RGD motif within the disintegrin domain, which is recognized by the integrins-␣5␤1 and -␣v␤3 (129).…”

Section: Other Adamsmentioning

confidence: 99%

ADAM-family metalloproteinases in lung inflammation: potential therapeutic targets

Dreymueller

Uhlig

Ludwig

2015

American Journal of Physiology-Lung Cellular and Molecular Phys

114

107

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-Acute and chronic lung inflammation is driven and controlled by several endogenous mediators that undergo proteolytic conversion from surface-expressed proteins to soluble variants by a disintegrin and metalloproteinase (ADAM)-family members. TNF and epidermal growth factor receptor ligands are just some of the many substrates by which these proteases regulate inflammatory or regenerative processes in the lung. ADAM10 and ADAM17 are the most prominent members of this protease family. They are constitutively expressed in most lung cells and, as recent research has shown, are the pivotal shedding enzymes mediating acute lung inflammation in a cell-specific manner. ADAM17 promotes endothelial and epithelial permeability, transendothelial leukocyte migration, and inflammatory mediator production by smooth muscle and epithelial cells. ADAM10 is critical for leukocyte migration and alveolar leukocyte recruitment. ADAM10 also promotes allergic asthma by driving B cell responses. Additionally, ADAM10 acts as a receptor for Staphylococcus aureus (S. aureus) ␣-toxin and is crucial for bacterial virulence. ADAM8, ADAM9, ADAM15, and ADAM33 are upregulated during acute or chronic lung inflammation, and recent functional or genetic analyses have linked them to disease development. Pharmacological inhibitors that allow us to locally or systemically target and differentiate ADAM-family members in the lung suppress acute and asthmatic inflammatory responses and S. aureus virulence. These promising results encourage further research to develop therapeutic strategies based on selected ADAMs. These studies need also to address the role of the ADAMs in repair and regeneration in the lung to identify further therapeutic opportunities and possible side effects. metalloproteinase; shedding; inflammation; edema; asthma ACUTE OR CHRONIC INFLAMMATION in the lung can lead to a fatal loss of lung functions. Acute inflammation resulting from infection, aspiration of gastric juice, or overventilation during intensive care is initially characterized by enhanced inflammatory mediator production, edema formation, and recruitment of innate immune cells with the risk of lung damage. Asthma, chronic obstructive pulmonary disease (COPD), and lung fibrosis are fatal chronic diseases that are driven by persistent inflammation with a lack of resolution and impaired tissue regeneration. Targeting the immune response is the underlying principle of many treatment strategies against these diseases (reviewed in Ref. 7).The cytokines, growth factors, receptors, and adhesion molecules that drive the inflammatory process are all under intensive scrutiny. Many of these molecules undergo functional modulation by limited proteolysis, bringing the participating proteases into the focus of attention. Among these proteases is a class of metalloproteinases that subdivides into the matrix metalloproteinases (MMP) and the families of a disintegrin and metalloproteinases (ADAM) and ADAM with trombospondin motif. MMPs have already been intensively investigated with resp...

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Characterization of the catalytic activity of the membrane-anchored metalloproteinase ADAM15 in cell-based assays

Cited by 47 publications

References 53 publications

Src Stimulates Fibroblast Growth Factor Receptor-2 Shedding by an ADAM15 Splice Variant Linked to Breast Cancer

Src Stimulates Fibroblast Growth Factor Receptor-2 Shedding by an ADAM15 Splice Variant Linked to Breast Cancer

Altered Bone Mechanics, Architecture and Composition in the Skeleton of TIMP-3-Deficient Mice

ADAM-family metalloproteinases in lung inflammation: potential therapeutic targets

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