EDTA/gelatin zymography method to identify C1s versus activated MMP-9 in plasma and immune complexes of patients with systemic lupus erythematosus.
Adolescent
Adult
Aged
Antigen-Antibody Complex
/ immunology
Edetic Acid
/ chemistry
Enzyme-Linked Immunosorbent Assay
/ methods
Female
Gelatin
/ chemistry
Gelatinases
/ metabolism
Humans
Lupus Erythematosus, Systemic
/ blood
Male
Matrix Metalloproteinase 2
/ blood
Matrix Metalloproteinase 9
/ blood
Middle Aged
Young Adult
EDTA/gelatin zymography
SLE marker
complement system subunit C1s
systemic lupus erythematosus
Journal
Journal of cellular and molecular medicine
ISSN: 1582-4934
Titre abrégé: J Cell Mol Med
Pays: England
ID NLM: 101083777
Informations de publication
Date de publication:
01 2019
01 2019
Historique:
received:
03
07
2018
revised:
11
09
2018
accepted:
20
09
2018
pubmed:
26
10
2018
medline:
16
4
2020
entrez:
26
10
2018
Statut:
ppublish
Résumé
Gelatin zymography analysis is a sensitive method and commonly used to characterize and quantify the presence of the gelatinases (MMP-2 and MMP-9) in biological samples. In human plasma samples from healthy controls and systemic lupus erythematosus (SLE) patients, we observed a gelatinolytic molecule at 80 kDa, suggestive for activated human MMP-9. However, by developing and using the EDTA/gelatin zymography method and after purification of the 80 kDa entity, we proved that this molecule was the C1s subunit of the complement system. The zymolytic capacity of C1s was validated and found to be enhanced, in the absence of calcium and in the presence of EDTA. Our findings indicate that for correct identification of gelatinolytic proteins in complex biological samples the use of EDTA/gelatin zymography for enzyme development is advised. In addition, by quantification of EDTA/gelatin zymography analysis and ELISA, we observed that the levels of C1s were higher in plasma and immune complexes of SLE patients than of healthy individuals. Therefore, our data imply that C1s may become a marker for the diagnosis of SLE.
Identifiants
pubmed: 30358100
doi: 10.1111/jcmm.13962
pmc: PMC6307758
doi:
Substances chimiques
Antigen-Antibody Complex
0
Gelatin
9000-70-8
Edetic Acid
9G34HU7RV0
Gelatinases
EC 3.4.24.-
Matrix Metalloproteinase 2
EC 3.4.24.24
Matrix Metalloproteinase 9
EC 3.4.24.35
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
576-585Informations de copyright
© 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.
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