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Tania Aaquist Ammitzbøll

Enzymatic and structural characterization of the major endopeptidase in the Venus flytrap digestion fluid

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Enzymatic and structural characterization of the major endopeptidase in the Venus flytrap digestion fluid. / Risor, Michael W; Thomsen, Line R; Sanggaard, Kristian W; Nielsen, Tania A; Thøgersen, Ida B; Lukassen, Marie V; Rossen, Litten; Garcia-Ferrer, Irene; Guevara, Tibisay; Scavenius, Carsten; Meinjohanns, Ernst; Gomis-Ruth, F Xavier; Enghild, Jan J.

In: Journal of Biological Chemistry, Vol. 291, 01.01.2016, p. 2271-2287.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Harvard

Risor, MW, Thomsen, LR, Sanggaard, KW, Nielsen, TA, Thøgersen, IB, Lukassen, MV, Rossen, L, Garcia-Ferrer, I, Guevara, T, Scavenius, C, Meinjohanns, E, Gomis-Ruth, FX & Enghild, JJ 2016, 'Enzymatic and structural characterization of the major endopeptidase in the Venus flytrap digestion fluid', Journal of Biological Chemistry, vol. 291, pp. 2271-2287. https://doi.org/10.1074/jbc.M115.672550

APA

Risor, M. W., Thomsen, L. R., Sanggaard, K. W., Nielsen, T. A., Thøgersen, I. B., Lukassen, M. V., Rossen, L., Garcia-Ferrer, I., Guevara, T., Scavenius, C., Meinjohanns, E., Gomis-Ruth, F. X., & Enghild, J. J. (2016). Enzymatic and structural characterization of the major endopeptidase in the Venus flytrap digestion fluid. Journal of Biological Chemistry, 291, 2271-2287. https://doi.org/10.1074/jbc.M115.672550

CBE

Risor MW, Thomsen LR, Sanggaard KW, Nielsen TA, Thøgersen IB, Lukassen MV, Rossen L, Garcia-Ferrer I, Guevara T, Scavenius C, Meinjohanns E, Gomis-Ruth FX, Enghild JJ. 2016. Enzymatic and structural characterization of the major endopeptidase in the Venus flytrap digestion fluid. Journal of Biological Chemistry. 291:2271-2287. https://doi.org/10.1074/jbc.M115.672550

MLA

Vancouver

Risor MW, Thomsen LR, Sanggaard KW, Nielsen TA, Thøgersen IB, Lukassen MV et al. Enzymatic and structural characterization of the major endopeptidase in the Venus flytrap digestion fluid. Journal of Biological Chemistry. 2016 Jan 1;291:2271-2287. https://doi.org/10.1074/jbc.M115.672550

Author

Risor, Michael W ; Thomsen, Line R ; Sanggaard, Kristian W ; Nielsen, Tania A ; Thøgersen, Ida B ; Lukassen, Marie V ; Rossen, Litten ; Garcia-Ferrer, Irene ; Guevara, Tibisay ; Scavenius, Carsten ; Meinjohanns, Ernst ; Gomis-Ruth, F Xavier ; Enghild, Jan J. / Enzymatic and structural characterization of the major endopeptidase in the Venus flytrap digestion fluid. In: Journal of Biological Chemistry. 2016 ; Vol. 291. pp. 2271-2287.

Bibtex

@article{6fc3a3f3c9984616b1c6216dd86a7e64,
title = "Enzymatic and structural characterization of the major endopeptidase in the Venus flytrap digestion fluid",
abstract = "Carnivorous plants primarily use aspartic proteases during digestion of captured prey. In contrast, the major endopeptidases in the digestive fluid of the Venus flytrap (Dionaea muscipula) are cysteine proteases (dionain-1 to -4). Here, we present the crystal structure of mature dionain-1 in covalent complex with inhibitor E-64 at 1.5 A resolution. The enzyme exhibits an overall protein fold reminiscent of other plant cysteine proteases. The inactive glycosylated pro-form undergoes autoprocessing and self-activation, optimally at the physiologically relevant pH value of 3.6, at which the protective effect of the pro-domain is lost. The mature enzyme was able to efficiently degrade a Drosophila fly protein extract at pH 4 showing high activity against the abundant Lys- and Arg-rich protein, myosin. The substrate specificity of dionain-1 was largely similar to that of papain with preference for hydrophobic and aliphatic residues in subsite S2 and for positively charged residues in S1. A tentative structure of the pro-domain was obtained by homology modeling and suggested that a pro-peptide Lys residue intrudes the S2 pocket which is more spacious than in papain. This study provides the first analysis of a cysteine protease from the digestive fluid of a carnivorous plant and confirms the close relationship between carnivorous action and plant defense mechanisms.",
author = "Risor, {Michael W} and Thomsen, {Line R} and Sanggaard, {Kristian W} and Nielsen, {Tania A} and Th{\o}gersen, {Ida B} and Lukassen, {Marie V} and Litten Rossen and Irene Garcia-Ferrer and Tibisay Guevara and Carsten Scavenius and Ernst Meinjohanns and Gomis-Ruth, {F Xavier} and Enghild, {Jan J}",
note = "Copyright {\textcopyright} 2015, The American Society for Biochemistry and Molecular Biology.",
year = "2016",
month = jan,
day = "1",
doi = "10.1074/jbc.M115.672550",
language = "English",
volume = "291",
pages = "2271--2287",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology, Inc.",

}

RIS

TY - JOUR

T1 - Enzymatic and structural characterization of the major endopeptidase in the Venus flytrap digestion fluid

AU - Risor, Michael W

AU - Thomsen, Line R

AU - Sanggaard, Kristian W

AU - Nielsen, Tania A

AU - Thøgersen, Ida B

AU - Lukassen, Marie V

AU - Rossen, Litten

AU - Garcia-Ferrer, Irene

AU - Guevara, Tibisay

AU - Scavenius, Carsten

AU - Meinjohanns, Ernst

AU - Gomis-Ruth, F Xavier

AU - Enghild, Jan J

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

PY - 2016/1/1

Y1 - 2016/1/1

N2 - Carnivorous plants primarily use aspartic proteases during digestion of captured prey. In contrast, the major endopeptidases in the digestive fluid of the Venus flytrap (Dionaea muscipula) are cysteine proteases (dionain-1 to -4). Here, we present the crystal structure of mature dionain-1 in covalent complex with inhibitor E-64 at 1.5 A resolution. The enzyme exhibits an overall protein fold reminiscent of other plant cysteine proteases. The inactive glycosylated pro-form undergoes autoprocessing and self-activation, optimally at the physiologically relevant pH value of 3.6, at which the protective effect of the pro-domain is lost. The mature enzyme was able to efficiently degrade a Drosophila fly protein extract at pH 4 showing high activity against the abundant Lys- and Arg-rich protein, myosin. The substrate specificity of dionain-1 was largely similar to that of papain with preference for hydrophobic and aliphatic residues in subsite S2 and for positively charged residues in S1. A tentative structure of the pro-domain was obtained by homology modeling and suggested that a pro-peptide Lys residue intrudes the S2 pocket which is more spacious than in papain. This study provides the first analysis of a cysteine protease from the digestive fluid of a carnivorous plant and confirms the close relationship between carnivorous action and plant defense mechanisms.

AB - Carnivorous plants primarily use aspartic proteases during digestion of captured prey. In contrast, the major endopeptidases in the digestive fluid of the Venus flytrap (Dionaea muscipula) are cysteine proteases (dionain-1 to -4). Here, we present the crystal structure of mature dionain-1 in covalent complex with inhibitor E-64 at 1.5 A resolution. The enzyme exhibits an overall protein fold reminiscent of other plant cysteine proteases. The inactive glycosylated pro-form undergoes autoprocessing and self-activation, optimally at the physiologically relevant pH value of 3.6, at which the protective effect of the pro-domain is lost. The mature enzyme was able to efficiently degrade a Drosophila fly protein extract at pH 4 showing high activity against the abundant Lys- and Arg-rich protein, myosin. The substrate specificity of dionain-1 was largely similar to that of papain with preference for hydrophobic and aliphatic residues in subsite S2 and for positively charged residues in S1. A tentative structure of the pro-domain was obtained by homology modeling and suggested that a pro-peptide Lys residue intrudes the S2 pocket which is more spacious than in papain. This study provides the first analysis of a cysteine protease from the digestive fluid of a carnivorous plant and confirms the close relationship between carnivorous action and plant defense mechanisms.

U2 - 10.1074/jbc.M115.672550

DO - 10.1074/jbc.M115.672550

M3 - Journal article

C2 - 26627834

VL - 291

SP - 2271

EP - 2287

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

ER -