Substrate specificity and inducibility of TACE (tumour necrosis factor α-converting enzyme) revisited: the Ala-Val preference, and induced intrinsic activity

Abstract

Tumour necrosis factor α (TNFα)-converting enzyme (TACE/ADAM-17, where ADAM stands for a disintegrin and metalloproteinase) releases from the cell surface the extracellular domains of TNF and several other proteins. Previous studies have found that, while purified TACE preferentially cleaves peptides representing the processing sites in TNF and transforming growth factor α, the cellular enzyme nonetheless also sheds proteins with divergent cleavage sites very efficiently. More recent work, identifying the cleavage site in the p75 TNF receptor, quantifying the susceptibility of additional peptides to cleavage by TACE and identifying additional protein substrates, underlines the complexity of TACE-substrate interactions. In addition to substrate specificity, the mechanism underlying the increased rate of shedding caused by agents that activate cells remains poorly understood. Recent work in this area, utilizing a peptide substrate as a probe for cellular TACE activity, indicates that the intrinsic activity of the enzyme is somehow increased.

Nectin-4, a New Serological Breast Cancer Marker, Is a Substrate for Tumor Necrosis Factor-α-converting Enzyme (TACE)/ADAM-17

“…In vitro, Nectin-4 cleavage by recombinant TACE results in the generation of a soluble form with a slightly higher molecular mass (data not shown). Differences in TACE cleavage position have been already reported in vitro using recombinant TACE and may explain this difference (29). The physiological inhibitor of TACE, TIMP-3, inhibits Nectin-4 cleavage.…”

Microparticles of Human Atherosclerotic Plaques Enhance the Shedding of the Tumor Necrosis Factor-α Converting Enzyme/ADAM17 Substrates, Tumor Necrosis Factor and Tumor Necrosis Factor Receptor-1

“…19 MPs (10 l) were suspended in 100 l of the final activity buffer (25 mmol/L Tris/HCl, pH 8.0, containing 2.5 mol/L ZnCl 2 ). Fluorogenic peptide I (substrate of MMP-1, -2, -7, -8, -9, -12, -13, -14, -15, and -16), peptide II (substrate of MMP-3 and -10) (R&D Systems), and peptide III were diluted in the activity buffer at the final concentration of 10 mol/L.…”

Activity of ADAM17 (a Disintegrin and Metalloprotease 17) Is Regulated by Its Noncatalytic Domains and Secondary Structure of its Substrates

“…For example, it was observed that ADAM17 failed to cleave several full-length proteins in biochemical assays but was able to cleave peptides based on their cleavage sites (reviewed by Roy Black in Ref. 38). Among the full-length proteins that exhibited this behavior were pro-TGF␣ (35), L-selectin, p55 TNFR, and p75 TNFR (38), all validated physiological substrates of ADAM17.…”

“…38). Among the full-length proteins that exhibited this behavior were pro-TGF␣ (35), L-selectin, p55 TNFR, and p75 TNFR (38), all validated physiological substrates of ADAM17. Moreover, the lack of suitable methodologies (long and expensive production process, sensitivity issues for HPLC-based methods, stability issues for fluorescamine labeling approach, etc.)…”