Document Detail


Functional analyses of complement convertases using C3 and C5-depleted sera.
MedLine Citation:
PMID:  23071769     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
C3 and C5 convertases are central stages of the complement cascade since they converge the different initiation pathways, augment complement activation by an amplification loop and lead to a common terminal pathway resulting in the formation of the membrane attack complex. Several complement inhibitors attenuate convertase formation and/or accelerate dissociation of convertase complexes. Functional assays used to study these processes are often performed using purified complement components, from which enzymatic complexes are reconstituted on the surface of erythrocytes or artificial matrices. This strategy enables identification of individual interactions between convertase components and putative regulators but carries an inherent risk of detecting non-physiological interactions that would not occur in a milieu of whole serum. Here we describe a novel, alternative method based on C3 or C5-depleted sera, which support activation of the complement cascade up to the desired stages of convertases. This approach allows fast and simple assessment of the influence of putative regulators on convertase formation and stability. As an example of practical utility of the assay, we performed studies on thioredoxin-1 in order to clarify the mechanism of its influence on complement convertases.
Authors:
Marcin Okroj; Emelie Holmquist; Ben C King; Anna M Blom
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-10-10
Journal Detail:
Title:  PloS one     Volume:  7     ISSN:  1932-6203     ISO Abbreviation:  PLoS ONE     Publication Date:  2012  
Date Detail:
Created Date:  2012-10-16     Completed Date:  2013-04-04     Revised Date:  2013-07-11    
Medline Journal Info:
Nlm Unique ID:  101285081     Medline TA:  PLoS One     Country:  United States    
Other Details:
Languages:  eng     Pagination:  e47245     Citation Subset:  IM    
Affiliation:
Department of Laboratory Medicine, Lund University, The Wallenberg Laboratory, Malmö, Sweden.
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MeSH Terms
Descriptor/Qualifier:
Chromatography, Affinity
Complement Activation / physiology*
Complement C3-C5 Convertases / physiology*
Enzyme Assays / methods*
Humans
Immune Sera / chemistry
Thioredoxins / chemistry,  metabolism
Chemical
Reg. No./Substance:
0/Immune Sera; 0/TXN protein, human; 52500-60-4/Thioredoxins; EC 3.4.21.-/Complement C3-C5 Convertases
Comments/Corrections

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