Recombinant Human TACE/ADAM17 Protein, CF

• Reactivity: Hu
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Human TACE/ADAM17 Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave a fluorogenic peptide substrate Mca-PLAQAV-Dpa-RSSSR-NH 2 (Catalog # ES003). The specific activity is >500 pmol/min/µg, as measured under the described conditions.
• Source: Spodoptera frugiperda, Sf 21 (baculovirus)-derived human TACE/ADAM17 protein Arg215-Asn671, with a C-terminal 6-His tag
• Accession #: P78536.1
• N-terminal Sequence: Arg215
• Structure / Form: Mature form. Recombinant Human TACE/ADAM17 may be prone to proteolytic cleavage at C-terminus. The poly-His tag may not be present in the preparation.
• Protein/Peptide Type: Recombinant Enzymes
• Gene: ADAM17
• Purity: >90%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 52 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 64 kDa, reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  6 months from date of receipt, -20 to -70 degreesC as supplied. 3 months, -20 to -70 degreesC under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in Tris, NaCl and Brij-35.
• Purity: >90%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile, deionized water.
• Assay Procedure:
  • Assay Buffer: 25 mM Tris, 2.5 μM ZnCl₂, 0.005% Brij-35 (w/v), pH 9.0 (note: It is extremely important that the assay solution does not contain salt (CaCl₂, NaCl, Na₂SO₄) because it inhibits TACE activity).
  • Recombinant Human TACE/ADAM17 (rhTACE) (Catalog # 930-ADB)
  • Substrate: MCA-Pro-Leu-Ala-Gln-Ala-Val-DPA-Arg-Ser-Ser-Ser-Arg-NH₂ (Catalog # ES003) , 2 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhTACE to 0.2 ng/µL in Assay Buffer.
  2. Dilute Substrate to 20 µM in Assay Buffer.
  3. In a plate load 50 µL of 0.2 ng/µL rhTACE and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 μL Assay Buffer and 50 µL Substrate.
  4. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively in kinetic mode for 5 minutes.
  5. Calculate specific activity:

  Specific Activity (pmol/min/µg) = (Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)) / (amount of enzyme (µg))

  *Adjusted for Substrate Blank
  **Derived using calibration standard MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).

  Per Well:
• rhTACE: 0.01 µg
• Substrate: 10 µM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human TACE/ADAM17 Protein, CF

• ADAM 17
• ADAM metallopeptidase domain 17
• ADAM metallopeptidase domain 18
• ADAM17
• ADAM18
• CD156b antigen
• CD156b
• CSVP
• disintegrin and metalloproteinase domain-containing protein 17
• EC 3.4.24.86
• MGC71942
• Snake venom-like protease
• TACE
• TACEcSVP
• TNF-alpha convertase
• TNF-alpha converting enzyme
• TNF-alpha-converting enzyme
• tumor necrosis factor, alpha, converting enzyme

Background

TACE is a member of the ADAM family that contains A Disintegrin And Metalloprotease-like domain. Like other membrane-anchored ADAMs, TACE consists of a pro domain with a cysteine switch and furin cleavage sequence, a catalytic domain with the zinc-binding site and Met-turn expected for reprolysins, a disintegrin-like domain, a cysteine-rich domain, an EGF-like domain, a transmembrane domain, and the cytoplasmic domain. In addition to its ability to release the 17 kDa extracellular form of tumor necrosis factor-alpha (TNF-alpha ) from the 26 kDa membrane-anchored TNF-alpha , TACE also plays an essential role in shedding ectodomains from a variety of proteins such as L-Selectin, Transforming Growth Factor-alpha , Amyloid Protein Precursor, and Notch-1 receptor. TACE mRNA is present in virtually every tissue and TACE protein resides both on the cell surface and in the cell.

1. Black, R.A. and J.D. Becherer (1998) in Tumor Necrosis Factor alpha -Converting Enzyme. Barrett, A.J. et al. (eds): Handbook of Proteolytic Enzymes, San Diego: Academic Press, p. 1315.
2. Primakoff, P. and D.G. Myles (2000) Trends in Genetics 16:83.

Publications for TACE/ADAM17 (930-ADB)(31)

Publications using 930-ADB

• Levengood, MR;Carosino, CM;Zhang, X;Lucas, S;Ortiz, DJ;Westendorf, L;Chin, AP;Martin, AD;Wong, A;Hengel, SM;Sun, H;Zeng, W;Yumul, R;Dominguez, MM;Chen, Y;Zheng, JH;Karlsson, CAB;Trang, VH;Senter, PD;Gardai, SJ; Preclinical development of SGN-CD47M: Protease-activated antibody technology enables selective tumor targeting of the innate immune checkpoint receptor CD47 Molecular cancer therapeutics 2024-10-28 [PMID: 39463068] (Bioassay, N/A)
• Huang, J;Zhao, C;Zhang, S; Semaphorin 7A promotes endothelial permeability and inflammation via plexin C1 and integrin ?1 in Kawasaki disease BMC pediatrics 2024-04-27 [PMID: 38678170] (Bioassay, Human)
• N Zhukovsky, M Silvano, T Filloux, S Gonzalez, KH Krause Alpha-1 Antitrypsin Reduces Disease Progression in a Mouse Model of Charcot-Marie-Tooth Type 1A: A Role for Decreased Inflammation and ADAM-17 Inhibition International Journal of Molecular Sciences, 2022-07-03;23(13):. 2022-07-03 [PMID: 35806409] (Bioassay, Human)
• DH Shin, SH Kim, M Choi, YK Bae, C Han, BK Choi, SS Kim, JY Han Oncogenic KRAS promotes growth of lung cancer cells expressing SLC3A2-NRG1 fusion via ADAM17-mediated shedding of NRG1 Oncogene, 2021-11-06;0(0):. 2021-11-06 [PMID: 34743207] (Bioassay, Human)
• Y Xiang, L Liu, Y Wang, B Li, J Peng, D Feng ADAM17 promotes the invasion of hepatocellular carcinoma via upregulation MMP21 Cancer Cell Int, 2020-10-21;20(0):516. 2020-10-21 [PMID: 33100908] (Bioassay, Human)
• AP Carreca, VM Pravatà, M Markham, S Bonelli, G Murphy, H Nagase, L Troeberg, SD Scilabra TIMP-3 facilitates binding of target metalloproteinases to the endocytic receptor LRP-1 and promotes scavenging of MMP-1 Sci Rep, 2020-07-21;10(1):12067. 2020-07-21 [PMID: 32694578] (Surface Plasmon Resonance, Human)
• M Maekawa, H Tadaki, D Tomimoto, C Okuma, R Sano, Y Ishii, Y Katsuda, H Yoshiuchi, R Kakefuda, T Ohta, T Sasase A Novel TNF-alpha Converting Enzyme (TACE) Selective Inhibitor JTP-96193 Prevents Insulin Resistance in KK-Ay Type 2 Diabetic Mice and Diabetic Peripheral Neuropathy in Type 1 Diabetic Mice Biol. Pharm. Bull., 2019-01-01;42(11):1906-1912. 2019-01-01 [PMID: 31685773] (Bioassay, Human)
• A Bertran, D Khomiak, A Konopka, E Rejmak, E Bulska, J Seco, L Kaczmarek, T Tarragó, R Prades Design and synthesis of selective and blood-brain barrier-permeable hydroxamate-based gelatinase inhibitors Bioorg. Chem., 2019-10-18;0(0):103365. 2019-10-18 [PMID: 31676116] (Bioassay, Human)
• A Ikeda, S Shahid, BR Blumberg, M Suzuki, JD Bartlett ADAM10 is Expressed by Ameloblasts, Cleaves the RELT TNF Receptor Extracellular Domain and Facilitates Enamel Development Sci Rep, 2019-10-01;9(1):14086. 2019-10-01 [PMID: 31575895] (Bioassay, Mouse)
• E Vezzoli, I Caron, F Talpo, D Besusso, P Conforti, E Battaglia, E Sogne, A Falqui, L Petricca, M Verani, P Martufi, A Caricasole, A Bresciani, O Cecchetti, P Rivetti di, G Sancini, O Riess, H Nguyen, L Seipold, P Saftig, G Biella, E Cattaneo, C Zuccato Inhibiting pathologically active ADAM10 rescues synaptic and cognitive decline in Huntington&#039;s disease J. Clin. Invest., 2019-05-06;130(0):. 2019-05-06 [PMID: 31063986] (Bioassay, Human)
• Y Ye, J Toczek, K Gona, HY Kim, J Han, M Razavian, R Golestani, J Zhang, TL Wu, M Ghosh, JJ Jung, MM Sadeghi Novel Arginine-containing Macrocyclic MMP Inhibitors: Synthesis, 99mTc-labeling, and Evaluation Sci Rep, 2018-08-03;8(1):11647. 2018-08-03 [PMID: 30076321] (Enzyme Assay, Human)
• T Yoneyama, M Gorry, A Sobo-Vujan, Y Lin, L Vujanovic, A Gaither-Da, ML Moss, MA Miller, LG Griffith, DA Lauffenbur, LP Stabile, J Herman, NL Vujanovic ADAM10 Sheddase Activity is a Potential Lung-Cancer Biomarker J Cancer, 2018-06-23;9(14):2559-2570. 2018-06-23 [PMID: 30026855] (Bioassay)
• JB Schaal, T Maretzky, DQ Tran, PA Tran, P Tongaonkar, CP Blobel, AJ Ouellette, ME Selsted Macrocyclic ?-defensins suppress tumor necrosis factor-? (TNF-?) shedding by inhibition of TNF-? converting enzyme J. Biol. Chem., 2018-01-09;0(0):. 2018-01-09 [PMID: 29317500] (Bioassay)
• Niacin-mediated Tace activation ameliorates CMT neuropathies with focal hypermyelination EMBO Mol Med, 2016-12-01;0(0):. 2016-12-01 [PMID: 27799291] (Bioassay, Mouse)
• Mira Jun The Identification of Biochanin A as a Potent and Selective ?-Site App-Cleaving Enzyme 1 (Bace1) Inhibitor Nutrients, 2016-10-14;8(10):. 2016-10-14 [PMID: 27754406] (Enzyme Assay)
• Mechanistic insights into a TIMP3-sensitive pathway constitutively engaged in the regulation of cerebral hemodynamics Elife, 2016-08-01;5(0):. 2016-08-01 [PMID: 27476853] (In Vivo, Mouse)
• Fibulin-1 Binds to Fibroblast Growth Factor 8 with High Affinity: Effects on Embryo Survival J Biol Chem, 2016-07-08;0(0):. 2016-07-08 [PMID: 27402846] (Bioassay, Human)
• Schlomann U, Koller G, Conrad C, Ferdous T, Golfi P, Garcia A, Hofling S, Parsons M, Costa P, Soper R, Bossard M, Hagemann T, Roshani R, Sewald N, Ketchem R, Moss M, Rasmussen F, Miller M, Lauffenburger D, Tuveson D, Nimsky C, Bartsch J ADAM8 as a drug target in pancreatic cancer. Nat Commun, 2015-01-28;6(0):6175. 2015-01-28 [PMID: 25629724] (Bioassay, Human)
• Lee E, Han J, Woo M, Shin J, Park E, Kang J, Moon P, Baek M, Son W, Ko Y, Choi J, Kim H Matrix metalloproteinase-8 plays a pivotal role in neuroinflammation by modulating TNF-alpha activation. J Immunol, 2014-07-21;193(5):2384-93. 2014-07-21 [PMID: 25049354] (Enzyme Assay)
• Lajoie L, Congy-Jolivet N, Bolzec A, Gouilleux-Gruart V, Sicard E, Sung H, Peiretti F, Moreau T, Vie H, Clemenceau B, Thibault G ADAM17-mediated shedding of FcgammaRIIIA on human NK cells: identification of the cleavage site and relationship with activation. J Immunol, 2013-12-13;192(2):741-51. 2013-12-13 [PMID: 24337742] (Bioassay, Human)
• McDonald A, Dibble J, Evans E, Millhauser G A new paradigm for enzymatic control of alpha-cleavage and beta-cleavage of the prion protein. J Biol Chem, 2013-11-18;289(2):803-13. 2013-11-18 [PMID: 24247244] (Enzyme Assay, Bacteria - E. Coli)
• Hayes C, Dey D, Palavicini J, Wang H, Patkar K, Minond D, Nefzi A, Lakshmana M Striking reduction of amyloid plaque burden in an Alzheimer&#039;s mouse model after chronic administration of carmustine. BMC Med, 2013-03-26;11(0):81. 2013-03-26 [PMID: 23531149] (Inhibition, Hamster)
• Moreira-Tabaka H, Peluso J, Vonesch JL, Hentsch D, Kessler P, Reimund JM, Dumont S, Muller CD Unlike for human monocytes after LPS activation, release of TNF-α by THP-1 cells is produced by a TACE catalytically different from constitutive TACE. PLoS ONE, 2012-03-30;7(3):e34184. 2012-03-30 [PMID: 22479555] (Enzyme Assay, Human)
• Gooz P, Dang Y, Higashiyama S, Twal WO, Haycraft CJ, Gooz M A disintegrin and metalloenzyme (ADAM) 17 activation is regulated by α5β1 integrin in kidney mesangial cells. PLoS ONE, 2012-03-08;7(3):e33350. 2012-03-08 [PMID: 22413019] (Binding Assay, Enzyme Assay, Human)
• Koenen RR, Pruessmeyer J, Soehnlein O, Fraemohs L, Zernecke A, Schwarz N, Reiss K, Sarabi A, Lindbom L, Hackeng TM, Weber C, Ludwig A Regulated release and functional modulation of junctional adhesion molecule A by disintegrin metalloproteinases. Blood, 2009-03-03;113(19):4799-809. 2009-03-03 [PMID: 19258599] (Bioassay, Enzyme Assay, Human)
• Moss ML, Bomar M, Liu Q, Sage H, Dempsey P, Lenhart PM, Gillispie PA, Stoeck A, Wildeboer D, Bartsch JW, Palmisano R, Zhou P The ADAM10 prodomain is a specific inhibitor of ADAM10 proteolytic activity and inhibits cellular shedding events. J. Biol. Chem., 2007-09-25;282(49):35712-21. 2007-09-25 [PMID: 17895248] (Bioassay, Human)
• Canault M, Leroyer AS, Peiretti F, Leseche G, Tedgui A, Bonardo B, Alessi MC, Boulanger CM, Nalbone G Microparticles of human atherosclerotic plaques enhance the shedding of the tumor necrosis factor-alpha converting enzyme/ADAM17 substrates, tumor necrosis factor and tumor necrosis factor receptor-1. Am. J. Pathol., 2007-09-14;171(5):1713-23. 2007-09-14 [PMID: 17872973] (Bioassay, Human)
• Tellier E, Rebsomen L The shedding activity of ADAM17 is sequestered in lipid rafts. Exp. Cell Res., 2006-09-05;312(20):3969-80. 2006-09-05 [PMID: 17010968] (Bioassay, Human)
• Nishi E, Hiraoka Y, Yoshida K, Okawa K, Kita T Nardilysin enhances ectodomain shedding of heparin-binding epidermal growth factor-like growth factor through activation of tumor necrosis factor-alpha-converting enzyme. J. Biol. Chem., 2006-08-21;281(41):31164-72. 2006-08-21 [PMID: 16923819] (Bioassay, Human)
• Hinkle CL, Sunnarborg SW, Loiselle D, Parker CE, Stevenson M, Russell WE, Lee DC Selective roles for tumor necrosis factor alpha-converting enzyme/ADAM17 in the shedding of the epidermal growth factor receptor ligand family: the juxtamembrane stalk determines cleavage efficiency. J. Biol. Chem., 2004-04-05;279(23):24179-88. 2004-04-05 [PMID: 15066986] (Enzyme Assay, Mouse)
• von Tresckow B, Kallen KJ, von Strandmann EP, Borchmann P, Lange H, Engert A, Hansen HP Depletion of cellular cholesterol and lipid rafts increases shedding of CD30. J. Immunol., 2004-04-01;172(7):4324-31. 2004-04-01 [PMID: 15034047] (Western Blot)

Bioinformatics

• Gene Symbol: ADAM17
• Uniprot:
  • P78536.1()