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Jens Christian Jensenius

Biological variations of MASP-3 and MAp44, two splice products of the MASP1 gene involved in regulation of the complement system

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Biological variations of MASP-3 and MAp44, two splice products of the MASP1 gene involved in regulation of the complement system. / Degn, Søren E; Jensen, Lisbeth; Gál, Péter; Dobó, József; Holmvad, Steffen H; Jensenius, Jens C; Thiel, Steffen.

In: Journal of Immunological Methods, Vol. 361, No. 1-2, 30.09.2010, p. 37-50.

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Degn, Søren E ; Jensen, Lisbeth ; Gál, Péter ; Dobó, József ; Holmvad, Steffen H ; Jensenius, Jens C ; Thiel, Steffen. / Biological variations of MASP-3 and MAp44, two splice products of the MASP1 gene involved in regulation of the complement system. In: Journal of Immunological Methods. 2010 ; Vol. 361, No. 1-2. pp. 37-50.

Bibtex

@article{44b5c23e93be4dbb8c50b3e805bb0265,
title = "Biological variations of MASP-3 and MAp44, two splice products of the MASP1 gene involved in regulation of the complement system",
abstract = "The lectin pathway of complement is part of the innate immune system. The complement-activating pattern-recognition molecules (for which we suggest the abbreviation CAPREMs) mannan-binding lectin (MBL) and the three ficolins (H-, L- and M-ficolin) circulate in complexes with MBL-associated serine proteases (MASP-1, -2 and -3) and two additional proteins (MAp19 and MAp44, also termed sMAP and MAP-1, respectively). When MBL or ficolins recognize a microorganism or altered self components, activation of the MASPs ensues, leading to the activation of the complement system. MASP-1, MASP-3 and MAp44 are all three encoded by the MASP1 gene. MASP-1 and -3 share five domains (constituting the so-called A-chain), but have unique protease domains (B-chains). MAp44 shares the first four domains with MASP-1 and MASP-3, followed by 17 unique C-terminal amino acid residues. Thus, assays for the protease domain of MASP-3 and for the 17 C-terminal amino acids of MAp44 are required to measure these proteins specifically and here we present such assays for MASP-3 and MAp44. MASP-3 was captured with a monoclonal antibody (5F5) reacting with a common domain of the three proteins (CCP1) and the assay was developed with a monoclonal antibody (38.12.3) specific for the C-terminal part of the MASP-3 protease domain. MAp44 was captured with a monoclonal antibody (2D5) reacting with the C-terminus of MAp44 followed by assay development with a monoclonal anti-CCP1 antibody (4H2). Using Superose 6 gel permeation chromatography of serum, MASP-3 and MAp44 were found in complexes, which eluted in positions corresponding to 600-800 kDa and 500-700 kDa, respectively. The level of MASP-3 in donor sera (N=200) was log-normally distributed with a median value of 5.0 μg/ml (range: 1.8-10.6 μg/ml), and the corresponding value for MAp44, also log-normally distributed, was 1.7 μg/ml (range: 0.8-3.2 μg/ml). For MASP-3, the inter-assay coefficients of variation of low, intermediate and high level internal controls were 4.9%, 6.9% and 3.9% (N=12). For MAp44, the corresponding inter-assay CVs were 7.6%, 6.2%, and 7.0% (N=12). MASP-3 levels were low at birth and reached adult levels within the first 6 months, whereas MAp44 levels fell slightly during the first 6 months. Concomitant with the acute phase response in patients undergoing major surgery, levels of both proteins fell slightly over 1-2 days, but whereas MASP-3 recovered to baseline values over another 2 days, MAp44 only reached baseline values at around day 30. Thus, neither of the two proteins behaves as a classical acute phase protein.",
keywords = "Adaptor Proteins, Signal Transducing, Adult, Amino Acid Sequence, Antibodies, Monoclonal, Apoptosis Regulatory Proteins, Blotting, Western, Complement Activation, Complement System Proteins, Humans, Infant, Newborn, Mannose-Binding Protein-Associated Serine Proteases, Molecular Sequence Data",
author = "Degn, {S{\o}ren E} and Lisbeth Jensen and P{\'e}ter G{\'a}l and J{\'o}zsef Dob{\'o} and Holmvad, {Steffen H} and Jensenius, {Jens C} and Steffen Thiel",
note = "Copyright {\textcopyright} 2010 Elsevier B.V. All rights reserved.",
year = "2010",
month = sep,
day = "30",
doi = "10.1016/j.jim.2010.07.006",
language = "English",
volume = "361",
pages = "37--50",
journal = "Journal of Immunological Methods",
issn = "0022-1759",
publisher = "Elsevier BV",
number = "1-2",

}

RIS

TY - JOUR

T1 - Biological variations of MASP-3 and MAp44, two splice products of the MASP1 gene involved in regulation of the complement system

AU - Degn, Søren E

AU - Jensen, Lisbeth

AU - Gál, Péter

AU - Dobó, József

AU - Holmvad, Steffen H

AU - Jensenius, Jens C

AU - Thiel, Steffen

N1 - Copyright © 2010 Elsevier B.V. All rights reserved.

PY - 2010/9/30

Y1 - 2010/9/30

N2 - The lectin pathway of complement is part of the innate immune system. The complement-activating pattern-recognition molecules (for which we suggest the abbreviation CAPREMs) mannan-binding lectin (MBL) and the three ficolins (H-, L- and M-ficolin) circulate in complexes with MBL-associated serine proteases (MASP-1, -2 and -3) and two additional proteins (MAp19 and MAp44, also termed sMAP and MAP-1, respectively). When MBL or ficolins recognize a microorganism or altered self components, activation of the MASPs ensues, leading to the activation of the complement system. MASP-1, MASP-3 and MAp44 are all three encoded by the MASP1 gene. MASP-1 and -3 share five domains (constituting the so-called A-chain), but have unique protease domains (B-chains). MAp44 shares the first four domains with MASP-1 and MASP-3, followed by 17 unique C-terminal amino acid residues. Thus, assays for the protease domain of MASP-3 and for the 17 C-terminal amino acids of MAp44 are required to measure these proteins specifically and here we present such assays for MASP-3 and MAp44. MASP-3 was captured with a monoclonal antibody (5F5) reacting with a common domain of the three proteins (CCP1) and the assay was developed with a monoclonal antibody (38.12.3) specific for the C-terminal part of the MASP-3 protease domain. MAp44 was captured with a monoclonal antibody (2D5) reacting with the C-terminus of MAp44 followed by assay development with a monoclonal anti-CCP1 antibody (4H2). Using Superose 6 gel permeation chromatography of serum, MASP-3 and MAp44 were found in complexes, which eluted in positions corresponding to 600-800 kDa and 500-700 kDa, respectively. The level of MASP-3 in donor sera (N=200) was log-normally distributed with a median value of 5.0 μg/ml (range: 1.8-10.6 μg/ml), and the corresponding value for MAp44, also log-normally distributed, was 1.7 μg/ml (range: 0.8-3.2 μg/ml). For MASP-3, the inter-assay coefficients of variation of low, intermediate and high level internal controls were 4.9%, 6.9% and 3.9% (N=12). For MAp44, the corresponding inter-assay CVs were 7.6%, 6.2%, and 7.0% (N=12). MASP-3 levels were low at birth and reached adult levels within the first 6 months, whereas MAp44 levels fell slightly during the first 6 months. Concomitant with the acute phase response in patients undergoing major surgery, levels of both proteins fell slightly over 1-2 days, but whereas MASP-3 recovered to baseline values over another 2 days, MAp44 only reached baseline values at around day 30. Thus, neither of the two proteins behaves as a classical acute phase protein.

AB - The lectin pathway of complement is part of the innate immune system. The complement-activating pattern-recognition molecules (for which we suggest the abbreviation CAPREMs) mannan-binding lectin (MBL) and the three ficolins (H-, L- and M-ficolin) circulate in complexes with MBL-associated serine proteases (MASP-1, -2 and -3) and two additional proteins (MAp19 and MAp44, also termed sMAP and MAP-1, respectively). When MBL or ficolins recognize a microorganism or altered self components, activation of the MASPs ensues, leading to the activation of the complement system. MASP-1, MASP-3 and MAp44 are all three encoded by the MASP1 gene. MASP-1 and -3 share five domains (constituting the so-called A-chain), but have unique protease domains (B-chains). MAp44 shares the first four domains with MASP-1 and MASP-3, followed by 17 unique C-terminal amino acid residues. Thus, assays for the protease domain of MASP-3 and for the 17 C-terminal amino acids of MAp44 are required to measure these proteins specifically and here we present such assays for MASP-3 and MAp44. MASP-3 was captured with a monoclonal antibody (5F5) reacting with a common domain of the three proteins (CCP1) and the assay was developed with a monoclonal antibody (38.12.3) specific for the C-terminal part of the MASP-3 protease domain. MAp44 was captured with a monoclonal antibody (2D5) reacting with the C-terminus of MAp44 followed by assay development with a monoclonal anti-CCP1 antibody (4H2). Using Superose 6 gel permeation chromatography of serum, MASP-3 and MAp44 were found in complexes, which eluted in positions corresponding to 600-800 kDa and 500-700 kDa, respectively. The level of MASP-3 in donor sera (N=200) was log-normally distributed with a median value of 5.0 μg/ml (range: 1.8-10.6 μg/ml), and the corresponding value for MAp44, also log-normally distributed, was 1.7 μg/ml (range: 0.8-3.2 μg/ml). For MASP-3, the inter-assay coefficients of variation of low, intermediate and high level internal controls were 4.9%, 6.9% and 3.9% (N=12). For MAp44, the corresponding inter-assay CVs were 7.6%, 6.2%, and 7.0% (N=12). MASP-3 levels were low at birth and reached adult levels within the first 6 months, whereas MAp44 levels fell slightly during the first 6 months. Concomitant with the acute phase response in patients undergoing major surgery, levels of both proteins fell slightly over 1-2 days, but whereas MASP-3 recovered to baseline values over another 2 days, MAp44 only reached baseline values at around day 30. Thus, neither of the two proteins behaves as a classical acute phase protein.

KW - Adaptor Proteins, Signal Transducing

KW - Adult

KW - Amino Acid Sequence

KW - Antibodies, Monoclonal

KW - Apoptosis Regulatory Proteins

KW - Blotting, Western

KW - Complement Activation

KW - Complement System Proteins

KW - Humans

KW - Infant, Newborn

KW - Mannose-Binding Protein-Associated Serine Proteases

KW - Molecular Sequence Data

U2 - 10.1016/j.jim.2010.07.006

DO - 10.1016/j.jim.2010.07.006

M3 - Journal article

C2 - 20673767

VL - 361

SP - 37

EP - 50

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

SN - 0022-1759

IS - 1-2

ER -