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Jens Christian Jensenius

Characterization of recombinant mannan-binding lectin-associated serine protease (MASP)-3 suggests an activation mechanism different from that of MASP-1 and MASP-2

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Characterization of recombinant mannan-binding lectin-associated serine protease (MASP)-3 suggests an activation mechanism different from that of MASP-1 and MASP-2. / Zundel, Stéphanie; Cseh, Sandor; Lacroix, Monique; Dahl, Mads R; Matsushita, Misao; Andrieu, Jean-Pierre; Schwaeble, Wilhelm J; Jensenius, Jens Christian; Fujita, Teizo; Arlaud, Gérard J; Thielens, Nicole M.

In: Journal of Immunology, Vol. 172, No. 7, 2004, p. 4342-50.

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

Harvard

Zundel, S, Cseh, S, Lacroix, M, Dahl, MR, Matsushita, M, Andrieu, J-P, Schwaeble, WJ, Jensenius, JC, Fujita, T, Arlaud, GJ & Thielens, NM 2004, 'Characterization of recombinant mannan-binding lectin-associated serine protease (MASP)-3 suggests an activation mechanism different from that of MASP-1 and MASP-2', Journal of Immunology, vol. 172, no. 7, pp. 4342-50.

APA

Zundel, S., Cseh, S., Lacroix, M., Dahl, M. R., Matsushita, M., Andrieu, J-P., Schwaeble, W. J., Jensenius, J. C., Fujita, T., Arlaud, G. J., & Thielens, N. M. (2004). Characterization of recombinant mannan-binding lectin-associated serine protease (MASP)-3 suggests an activation mechanism different from that of MASP-1 and MASP-2. Journal of Immunology, 172(7), 4342-50.

CBE

Zundel S, Cseh S, Lacroix M, Dahl MR, Matsushita M, Andrieu J-P, Schwaeble WJ, Jensenius JC, Fujita T, Arlaud GJ, Thielens NM. 2004. Characterization of recombinant mannan-binding lectin-associated serine protease (MASP)-3 suggests an activation mechanism different from that of MASP-1 and MASP-2. Journal of Immunology. 172(7):4342-50.

MLA

Vancouver

Author

Zundel, Stéphanie ; Cseh, Sandor ; Lacroix, Monique ; Dahl, Mads R ; Matsushita, Misao ; Andrieu, Jean-Pierre ; Schwaeble, Wilhelm J ; Jensenius, Jens Christian ; Fujita, Teizo ; Arlaud, Gérard J ; Thielens, Nicole M. / Characterization of recombinant mannan-binding lectin-associated serine protease (MASP)-3 suggests an activation mechanism different from that of MASP-1 and MASP-2. In: Journal of Immunology. 2004 ; Vol. 172, No. 7. pp. 4342-50.

Bibtex

@article{5f6c9c1bc7c246329104c72dca7b4d2a,
title = "Characterization of recombinant mannan-binding lectin-associated serine protease (MASP)-3 suggests an activation mechanism different from that of MASP-1 and MASP-2",
abstract = "Mannan-binding lectin (MBL)-associated serine proteases (MASP-1, -2, and -3) are homologous modular proteases that each associate with MBL and L- and H-ficolins, which are oligomeric serum lectins involved in innate immunity. To investigate its physicochemical, interaction, and enzymatic properties, human MASP-3 was expressed in insect cells. Ultracentrifugation analysis indicated that rMASP-3 sedimented as a homodimer (s(20,w) = 6.2 +/- 0.1 S) in the presence of Ca(2+), and as a monomer (s(20,w) = 4.6 +/- 0.1 S) in EDTA. As shown by surface plasmon resonance spectroscopy, it associated with both MBL (K(D) = 2.6 nM) and L-ficolin (K(D) = 7.2 nM). The protease was produced in a single-chain, proenzyme form, but underwent slow activation upon prolonged storage at 4 degrees C, resulting from cleavage at the Arg(430)-Ile(431) activation site. Activation was prevented in the presence of protease inhibitors iodoacetamide and 1,10-phenanthroline but was not abolished upon substitution of Ala for the active site Ser(645) of MASP-3, indicating extrinsic proteolysis. In contrast, the corresponding mutations Ser(627)-->Ala in MASP-1 and Ser(618)-->Ala in MASP-2 stabilized the latter in their proenzyme form. Likewise, the MASP-1 and MASP-2 mutants were each activated by their active counterparts, but MASP-3 S645A was not. Activated MASP-3 did not react with C1 inhibitor; had no activity on complement proteins C2, C4, and C3; and only cleaved the N-carboxybenzyloxyglycine-L-arginine thiobenzyl ester substrate to a significant extent. Based on these observations, it is postulated that MASP-3 activation and control involve mechanisms that are different from those of MASP-1 and -2.",
keywords = "Alanine, Amino Acid Substitution, Animals, Baculoviridae, Carrier Proteins, Complement Activation, Complement C1 Inactivator Proteins, Enzyme Activation, Humans, Lectins, Mannose-Binding Lectin, Mannose-Binding Lectins, Mannose-Binding Protein-Associated Serine Proteases, Recombinant Proteins, Serine, Serine Endopeptidases, Serpins, Spodoptera",
author = "St{\'e}phanie Zundel and Sandor Cseh and Monique Lacroix and Dahl, {Mads R} and Misao Matsushita and Jean-Pierre Andrieu and Schwaeble, {Wilhelm J} and Jensenius, {Jens Christian} and Teizo Fujita and Arlaud, {G{\'e}rard J} and Thielens, {Nicole M}",
year = "2004",
language = "English",
volume = "172",
pages = "4342--50",
journal = "Journal of Immunology",
issn = "0022-1767",
publisher = "American Association of Immunologists",
number = "7",

}

RIS

TY - JOUR

T1 - Characterization of recombinant mannan-binding lectin-associated serine protease (MASP)-3 suggests an activation mechanism different from that of MASP-1 and MASP-2

AU - Zundel, Stéphanie

AU - Cseh, Sandor

AU - Lacroix, Monique

AU - Dahl, Mads R

AU - Matsushita, Misao

AU - Andrieu, Jean-Pierre

AU - Schwaeble, Wilhelm J

AU - Jensenius, Jens Christian

AU - Fujita, Teizo

AU - Arlaud, Gérard J

AU - Thielens, Nicole M

PY - 2004

Y1 - 2004

N2 - Mannan-binding lectin (MBL)-associated serine proteases (MASP-1, -2, and -3) are homologous modular proteases that each associate with MBL and L- and H-ficolins, which are oligomeric serum lectins involved in innate immunity. To investigate its physicochemical, interaction, and enzymatic properties, human MASP-3 was expressed in insect cells. Ultracentrifugation analysis indicated that rMASP-3 sedimented as a homodimer (s(20,w) = 6.2 +/- 0.1 S) in the presence of Ca(2+), and as a monomer (s(20,w) = 4.6 +/- 0.1 S) in EDTA. As shown by surface plasmon resonance spectroscopy, it associated with both MBL (K(D) = 2.6 nM) and L-ficolin (K(D) = 7.2 nM). The protease was produced in a single-chain, proenzyme form, but underwent slow activation upon prolonged storage at 4 degrees C, resulting from cleavage at the Arg(430)-Ile(431) activation site. Activation was prevented in the presence of protease inhibitors iodoacetamide and 1,10-phenanthroline but was not abolished upon substitution of Ala for the active site Ser(645) of MASP-3, indicating extrinsic proteolysis. In contrast, the corresponding mutations Ser(627)-->Ala in MASP-1 and Ser(618)-->Ala in MASP-2 stabilized the latter in their proenzyme form. Likewise, the MASP-1 and MASP-2 mutants were each activated by their active counterparts, but MASP-3 S645A was not. Activated MASP-3 did not react with C1 inhibitor; had no activity on complement proteins C2, C4, and C3; and only cleaved the N-carboxybenzyloxyglycine-L-arginine thiobenzyl ester substrate to a significant extent. Based on these observations, it is postulated that MASP-3 activation and control involve mechanisms that are different from those of MASP-1 and -2.

AB - Mannan-binding lectin (MBL)-associated serine proteases (MASP-1, -2, and -3) are homologous modular proteases that each associate with MBL and L- and H-ficolins, which are oligomeric serum lectins involved in innate immunity. To investigate its physicochemical, interaction, and enzymatic properties, human MASP-3 was expressed in insect cells. Ultracentrifugation analysis indicated that rMASP-3 sedimented as a homodimer (s(20,w) = 6.2 +/- 0.1 S) in the presence of Ca(2+), and as a monomer (s(20,w) = 4.6 +/- 0.1 S) in EDTA. As shown by surface plasmon resonance spectroscopy, it associated with both MBL (K(D) = 2.6 nM) and L-ficolin (K(D) = 7.2 nM). The protease was produced in a single-chain, proenzyme form, but underwent slow activation upon prolonged storage at 4 degrees C, resulting from cleavage at the Arg(430)-Ile(431) activation site. Activation was prevented in the presence of protease inhibitors iodoacetamide and 1,10-phenanthroline but was not abolished upon substitution of Ala for the active site Ser(645) of MASP-3, indicating extrinsic proteolysis. In contrast, the corresponding mutations Ser(627)-->Ala in MASP-1 and Ser(618)-->Ala in MASP-2 stabilized the latter in their proenzyme form. Likewise, the MASP-1 and MASP-2 mutants were each activated by their active counterparts, but MASP-3 S645A was not. Activated MASP-3 did not react with C1 inhibitor; had no activity on complement proteins C2, C4, and C3; and only cleaved the N-carboxybenzyloxyglycine-L-arginine thiobenzyl ester substrate to a significant extent. Based on these observations, it is postulated that MASP-3 activation and control involve mechanisms that are different from those of MASP-1 and -2.

KW - Alanine

KW - Amino Acid Substitution

KW - Animals

KW - Baculoviridae

KW - Carrier Proteins

KW - Complement Activation

KW - Complement C1 Inactivator Proteins

KW - Enzyme Activation

KW - Humans

KW - Lectins

KW - Mannose-Binding Lectin

KW - Mannose-Binding Lectins

KW - Mannose-Binding Protein-Associated Serine Proteases

KW - Recombinant Proteins

KW - Serine

KW - Serine Endopeptidases

KW - Serpins

KW - Spodoptera

M3 - Journal article

C2 - 15034049

VL - 172

SP - 4342

EP - 4350

JO - Journal of Immunology

JF - Journal of Immunology

SN - 0022-1767

IS - 7

ER -