Glycosylation of a disintegrin and metalloprotease 17 affects its activity and inhibition
Open accessAbstract
ADAM17 (a disintegrin and metalloprotease 17) is believed to be a tractable target in various diseases, including cancer and rheumatoid arthritis; however, it is not known whether glycosylation of ADAM17 expressed in healthy cells differs from that found in diseased tissue and, if so, whether glycosylation affects inhibitor binding. We expressed human ADAM17 in mammalian and insect cells and compared their glycosylation, substrate kinetics, and inhibition profiles. We found that ADAM17 expressed in mammalian cells was more heavily glycosylated than its insect-expressed analog. To determine whether differential glycosylation modulates enzymatic activity, we performed kinetic studies with both ADAM17 analogs and various TNFα-based substrates. The mammalian form of ADAM17 exhibited 10- to 30-fold lower kcat values than the insect analog, while the KM was unaffected, suggesting that glycosylation of ADAM17 can potentially play a role in regulating enzyme activity in vivo. Finally, we tested ADAM17 forms for inhibition by several well-characterized inhibitors. Active-site zinc-binding small molecules did not exhibit differences between the two ADAM17 analogs, while a non-zinc-binding exosite inhibitor of ADAM17 showed significantly lower potency toward the mammalian-expressed analog. These results suggest that glycosylation of ADAM17 can affect cell signaling in disease and might provide opportunities for therapeutic intervention using exosite inhibitors.
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Amyloidogenic and non-amyloidogenic processing of APP is influenced not only by the biochemical properties of APP but also by the biochemical properties of the secretases.
“Amyloidogenic and non-amyloidogenic processing of APP is influence not only by the biochemical properties of APP but also by the biochemical properties of the secretases 21,24,26 .”
From Targeting Lysosomal Dysfunction to Alleviate Plaque Deposition in an Alzheimer Disease Model (2025) · cites this paper as [26]
The Introduction (page unavailable) reports that ADAM17’s catalytic properties depend on its expression system and that its glycosylation may influence substrate binding. This supports the general point that a protease’s biochemical properties can affect processing, but the paper does not examine APP or establish effects on its amyloidogenic and non-amyloidogenic pathways. (LLM, conf=0.83)
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Research works printed but not linked: 24
- [1]Murphy G The ADAMs: signalling scissors in the tumour microenvironment Nat Rev Cancer 8 2008 932 941 19005493
- [2]Endres K Anders A Kojro E Gilbert S Fahrenholz F Postina R Tumor necrosis factor-α converting enzyme is processed by proprotein-convertases to its mature form which is degraded upon phorbol ester stimulation European Journal of Biochemistry 270 2003 2386 2393 12755693
- [3]Moss ML Jin SLC Milla ME Burkhart W Carter HL Chen WJ Clay WC Didsbury JR Hassler D Hoffman CR Kost TA Lambert MH Leesnitzer MA McCauley P McGeehan G Mitchell J Moyer M Pahel G Rocque W Overton LK Schoenen F Seaton T Su JL Warner J Willard D Becherer JD Cloning of a disintegrin metalloproteinase that processes precursor tumour-necrosis factor-[alpha] Nature 385 1997 733 736 9034191
- [4]Primakoff P Myles DG The ADAM gene family: surface proteins with adhesion and protease activity Trends in Genetics 16 2000 83 87 10652535
- [5]Edwards DR Handsley MM Pennington CJ The ADAM metalloproteinases Molecular Aspects of Medicine 29 2008 258 289 18762209 PMC7112278
- [6]Reiss K Saftig P The “A Disintegrin And Metalloprotease” (ADAM) family of sheddases: Physiological and cellular functions Seminars in Cell & Developmental Biology 20 2009 126 137 19049889
- [7]Murphy G The ADAMs: signalling scissors in the tumour microenvironment Nat Rev Cancer 8 2008 929 941 19005493
- [8]Borrell-Pages M Rojo F Albanell J Baselga J Arribas J TACE is required for the activation of the EGFR by TGF-alpha in tumors EMBO J 22 2003 1114 1124 12606576 PMC150344
- [9]Condon TP Flournoy S Sawyer GJ Baker BF Kishimoto TK Bennett CF ADAM17 but Not ADAM10 Mediates Tumor Necrosis Factor-α and L-Selectin Shedding from Leukocyte Membranes ADAM17 but Not ADAM10 Mediates Tumor Necrosis Factor-α and L-Selectin Shedding from Leukocyte Membranes 11 2001 107 116 11334139
- [10]Georgiadis D Yiotakis A Specific targeting of metzincin family members with small-molecule inhibitors: progress toward a multifarious challenge Bioorg Med Chem 16 2008 8781 8794 18790648
- [11]Tape CJ Willems SH Dombernowsky SL Stanley PL Fogarasi M Ouwehand W McCafferty J Murphy G Cross-domain inhibition of TACE ectodomain Proceedings of the National Academy of Sciences 108 2011 5578 5583 PMC3078358 21415364
- [12]Minond D Cudic M Bionda N Giulianotti M Maida L Houghten RA Fields GB Discovery of Novel Inhibitors of A Disintegrin And Metalloprotease 17 (ADAM17) Using Glycosylated and Non-Glycosylated Substrates Journal of Biological Chemistry 2012 PMC3476313 22927435
- [13]Black RA Rauch CT Kozlosky CJ Peschon JJ Slack JL Wolfson MF Castner BJ Stocking KL Reddy P Srinivasan S Nelson N Boiani N Schooley KA Gerhart M Davis R Fitzner JN Johnson RS Paxton RJ March CJ Cerretti DP A metalloproteinase disintegrin that releases tumour-necrosis factor-alpha from cells Nature 385 1997 729 733 9034190
- [15]Rolain T Bernard E Beaussart A Degand H Courtin P Egge-Jacobsen W Bron PA Morsomme P Kleerebezem M Chapot-Chartier MP Dufrene YF Hols P O-glycosylation as a novel control mechanism of peptidoglycan hydrolase activity J Biol Chem 2013 PMC3829315 23760506
- [16]Liebminger E Grass J Altmann F Mach L Strasser R Characterizing the link between glycosylation state and enzymatic activity of the endo-beta1,4-glucanase KORRIGAN1 from Arabidopsis thaliana J Biol Chem 2013 PMC3829318 23782689
- [17]Vandooren J Van den Steen PE Opdenakker G Biochemistry and molecular biology of gelatinase B or matrix metalloproteinase-9 (MMP-9): The next decade Crit Rev Biochem Mol Biol 48 2013 222 272 23547785
- [18]Copeland RA Enzymes 2 Wiley-VCH, Inc 2000
- [19]Wei S Xie Z Filenova E Brew K Drosophila TIMP is a potent inhibitor of MMPs and TACE: similarities in structure and function to TIMP-3 Biochemistry 42 2003 12200 12207 14567681
- [20]Niu X Umland S Ingram R Beyer BM Liu YH Sun J Lundell D Orth P IK682, a tight binding inhibitor of TACE Arch Biochem Biophys 451 2006 43 50 16762314
- [21]Neumann U Kubota H Frei K Ganu V Leppert D Characterization of Mca-Lys-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2, a fluorogenic substrate with increased specificity constants for collagenases and tumor necrosis factor converting enzyme Anal Biochem 328 2004 166 173 15113693
- [22]Moss ML Rasmussen FH Fluorescent substrates for the proteinases ADAM17, ADAM10, ADAM8, and ADAM12 useful for high-throughput inhibitor screening Anal Biochem 366 2007 144 148 17548045
- [23]Moss ML Rasmussen FH Nudelman R Dempsey PJ Williams J Fluorescent substrates useful as high-throughput screening tools for ADAM9 Comb Chem High Throughput Screen 13 2010 358 365 20015014
- [24]Minond D Cudic M Bionda N Giulianotti M Maida L Houghten RA Fields GB Discovery of Novel Inhibitors of a Disintegrin and Metalloprotease 17 (ADAM17) Using Glycosylated and Non-glycosylated Substrates J Biol Chem 287 2012 36473 36487 22927435 PMC3476313
- [25]Stawikowska R Cudic M Giulianotti M Houghten RA Fields GB Minond D Activity of a disintegrin and metalloprotease 17 (ADAM17) is Regulated by its non-catalytic domains and secondary structure of its substrates J Biol Chem 2013 PMC3829370 23779109
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Targeting Lysosomal Dysfunction to Alleviate Plaque Deposition in an Alzheimer Disease Model
Fremuth, L. E., van de Vlekkert, D., Hu, H., Weesner, J. A., Annunziata, I., Yogalingam, G., & d’Azzo, A. (2025) · doi:10.1101/2025.04.28.651121 · Cited by 1 · see citing works
- Authors
- Chavaroche, A., Cudic, M., Giulianotti, M., Houghten, R. A., Fields, G. B., & Minond, D.
- Affiliations
- Torrey Pines Institute For Molecular Studies (US)
- Corresponding author
- Minond, Dmitriy
- Venue
- Analytical Biochemistry
- Publisher
- Elsevier BV
- Published
- 2014-03
- Volume
- 449
- Pages
- 68-75
- ISSN
- 0003-2697 (ISSN-L)
- PMC
- PMC4334441
- OpenAlex
- W2040799583
- Cited by
- 30
- References
- 0
- Citations by year
- 2026: 7 · 2025: 3 · 2024: 1 · 2023: 1 · 2022: 2 · 2021: 4 · 2020: 2 · 2019: 2 · 2018: 2 · 2017: 1 · 2016: 2 · 2015: 1 · 2014: 2
- FWCI
- 0.49
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Analytical Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institute on Drug Abuse (R03DA033985, DA033985, R01DA031370, DA031370)
- National Cancer Institute (CA098799, R01CA098799)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Protease and Inhibitor Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glycosylation and Glycoproteins Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
- Keywords
- ADAM17, glycosylation, TNF-alpha processing, enzyme kinetics, substrate specificity, signal transduction
- MeSH terms
- ADAM17 Protein, Amino Acid Sequence, Animals, Cloning, Molecular, Enzyme Activation, Enzyme Inhibitors, Glycosylation, Humans, Kinetics, Molecular Sequence Data, Recombinant Proteins, ADAM Proteins, HEK293 Cells
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