Evidence for Restricted Reactivity of ADAMDEC1 with Protein Substrates and Endogenous Inhibitors

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Evidence for Restricted Reactivity of ADAMDEC1 with Protein Substrates and Endogenous Inhibitors. / Lund, Jacob; Troeberg, Linda; Kjeldal, Henrik; Olsen, Ole H; Nagase, Hideaki; Sørensen, Esben Skipper; Stennicke, Henning R; Petersen, Helle H; Overgaard, Michael Toft.

I: Journal of Biological Chemistry, Bind 290, 06.03.2015, s. 6620-6629.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

Harvard

Lund, J, Troeberg, L, Kjeldal, H, Olsen, OH, Nagase, H, Sørensen, ES, Stennicke, HR, Petersen, HH & Overgaard, MT 2015, 'Evidence for Restricted Reactivity of ADAMDEC1 with Protein Substrates and Endogenous Inhibitors', Journal of Biological Chemistry, bind 290, s. 6620-6629. https://doi.org/10.1074/jbc.M114.601724

APA

Lund, J., Troeberg, L., Kjeldal, H., Olsen, O. H., Nagase, H., Sørensen, E. S., Stennicke, H. R., Petersen, H. H., & Overgaard, M. T. (2015). Evidence for Restricted Reactivity of ADAMDEC1 with Protein Substrates and Endogenous Inhibitors. Journal of Biological Chemistry, 290, 6620-6629. https://doi.org/10.1074/jbc.M114.601724

CBE

Lund J, Troeberg L, Kjeldal H, Olsen OH, Nagase H, Sørensen ES, Stennicke HR, Petersen HH, Overgaard MT. 2015. Evidence for Restricted Reactivity of ADAMDEC1 with Protein Substrates and Endogenous Inhibitors. Journal of Biological Chemistry. 290:6620-6629. https://doi.org/10.1074/jbc.M114.601724

MLA

Vancouver

Lund J, Troeberg L, Kjeldal H, Olsen OH, Nagase H, Sørensen ES o.a. Evidence for Restricted Reactivity of ADAMDEC1 with Protein Substrates and Endogenous Inhibitors. Journal of Biological Chemistry. 2015 mar 6;290:6620-6629. https://doi.org/10.1074/jbc.M114.601724

Author

Lund, Jacob ; Troeberg, Linda ; Kjeldal, Henrik ; Olsen, Ole H ; Nagase, Hideaki ; Sørensen, Esben Skipper ; Stennicke, Henning R ; Petersen, Helle H ; Overgaard, Michael Toft. / Evidence for Restricted Reactivity of ADAMDEC1 with Protein Substrates and Endogenous Inhibitors. I: Journal of Biological Chemistry. 2015 ; Bind 290. s. 6620-6629.

Bibtex

@article{2a9ada23c42640a2b991d0cd34c6f5ff,
title = "Evidence for Restricted Reactivity of ADAMDEC1 with Protein Substrates and Endogenous Inhibitors",
abstract = "ADAMDEC1 is a proteolytically active metzincin metalloprotease displaying rare active site architecture with a zinc-binding Asp residue (Asp362). We previously demonstrated that substitution of Asp362 for a His residue, thereby reconstituting the canonical metzincin zinc-binding environment with three His zinc-ligands, increases the proteolytic activity. The protease also has an atypically short domain structure with an odd number of Cys residues in the metalloprotease domain. Here, we investigated how these rare structural features in the ADAMDEC1 metalloprotease domain impact the proteolytic activity, substrate specificity, and the effect of inhibitors. We identified carboxymethylated transferrin (Cm-Tf) as a new ADAMDEC1 substrate and determined the primary and secondary cleavage sites, which suggests a strong preference for Leu in the P1' position. Cys392 present in human, but only partially conserved within sequenced ADAMDEC1 orthologs, was found to be unpaired, and substitution of Cys392 for a Ser increased the reactivity with α2-macroglobulin, but not with casein or Cm-Tf. Substitution of Asp362 for His resulted in a general increase in proteolytic activity and a change in substrate specificity was observed with Cm-Tf. ADAMDEC1 was inhibited by the small molecule inhibitor Batimastat, but not by tissue inhibitor of metalloproteases (TIMP)-1, TIMP-2 or the N-terminal inhibitory domain of TIMP-3 (N-TIMP-3). However, N-TIMP-3 displayed profound inhibitory activity against the Asp362His variants with a reconstituted consensus metzincin zinc-binding environment. We hypothesize that these unique features of ADAMDEC1 may have evolved to escape from inhibition by endogenous metalloprotease inhibitors.",
author = "Jacob Lund and Linda Troeberg and Henrik Kjeldal and Olsen, {Ole H} and Hideaki Nagase and S{\o}rensen, {Esben Skipper} and Stennicke, {Henning R} and Petersen, {Helle H} and Overgaard, {Michael Toft}",
note = "Copyright {\textcopyright} 2015, The American Society for Biochemistry and Molecular Biology.",
year = "2015",
month = mar,
day = "6",
doi = "10.1074/jbc.M114.601724",
language = "English",
volume = "290",
pages = "6620--6629",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology, Inc.",

}

RIS

TY - JOUR

T1 - Evidence for Restricted Reactivity of ADAMDEC1 with Protein Substrates and Endogenous Inhibitors

AU - Lund, Jacob

AU - Troeberg, Linda

AU - Kjeldal, Henrik

AU - Olsen, Ole H

AU - Nagase, Hideaki

AU - Sørensen, Esben Skipper

AU - Stennicke, Henning R

AU - Petersen, Helle H

AU - Overgaard, Michael Toft

N1 - Copyright © 2015, The American Society for Biochemistry and Molecular Biology.

PY - 2015/3/6

Y1 - 2015/3/6

N2 - ADAMDEC1 is a proteolytically active metzincin metalloprotease displaying rare active site architecture with a zinc-binding Asp residue (Asp362). We previously demonstrated that substitution of Asp362 for a His residue, thereby reconstituting the canonical metzincin zinc-binding environment with three His zinc-ligands, increases the proteolytic activity. The protease also has an atypically short domain structure with an odd number of Cys residues in the metalloprotease domain. Here, we investigated how these rare structural features in the ADAMDEC1 metalloprotease domain impact the proteolytic activity, substrate specificity, and the effect of inhibitors. We identified carboxymethylated transferrin (Cm-Tf) as a new ADAMDEC1 substrate and determined the primary and secondary cleavage sites, which suggests a strong preference for Leu in the P1' position. Cys392 present in human, but only partially conserved within sequenced ADAMDEC1 orthologs, was found to be unpaired, and substitution of Cys392 for a Ser increased the reactivity with α2-macroglobulin, but not with casein or Cm-Tf. Substitution of Asp362 for His resulted in a general increase in proteolytic activity and a change in substrate specificity was observed with Cm-Tf. ADAMDEC1 was inhibited by the small molecule inhibitor Batimastat, but not by tissue inhibitor of metalloproteases (TIMP)-1, TIMP-2 or the N-terminal inhibitory domain of TIMP-3 (N-TIMP-3). However, N-TIMP-3 displayed profound inhibitory activity against the Asp362His variants with a reconstituted consensus metzincin zinc-binding environment. We hypothesize that these unique features of ADAMDEC1 may have evolved to escape from inhibition by endogenous metalloprotease inhibitors.

AB - ADAMDEC1 is a proteolytically active metzincin metalloprotease displaying rare active site architecture with a zinc-binding Asp residue (Asp362). We previously demonstrated that substitution of Asp362 for a His residue, thereby reconstituting the canonical metzincin zinc-binding environment with three His zinc-ligands, increases the proteolytic activity. The protease also has an atypically short domain structure with an odd number of Cys residues in the metalloprotease domain. Here, we investigated how these rare structural features in the ADAMDEC1 metalloprotease domain impact the proteolytic activity, substrate specificity, and the effect of inhibitors. We identified carboxymethylated transferrin (Cm-Tf) as a new ADAMDEC1 substrate and determined the primary and secondary cleavage sites, which suggests a strong preference for Leu in the P1' position. Cys392 present in human, but only partially conserved within sequenced ADAMDEC1 orthologs, was found to be unpaired, and substitution of Cys392 for a Ser increased the reactivity with α2-macroglobulin, but not with casein or Cm-Tf. Substitution of Asp362 for His resulted in a general increase in proteolytic activity and a change in substrate specificity was observed with Cm-Tf. ADAMDEC1 was inhibited by the small molecule inhibitor Batimastat, but not by tissue inhibitor of metalloproteases (TIMP)-1, TIMP-2 or the N-terminal inhibitory domain of TIMP-3 (N-TIMP-3). However, N-TIMP-3 displayed profound inhibitory activity against the Asp362His variants with a reconstituted consensus metzincin zinc-binding environment. We hypothesize that these unique features of ADAMDEC1 may have evolved to escape from inhibition by endogenous metalloprotease inhibitors.

U2 - 10.1074/jbc.M114.601724

DO - 10.1074/jbc.M114.601724

M3 - Journal article

C2 - 25564618

VL - 290

SP - 6620

EP - 6629

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

ER -