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Functional analyses of complement convertases using c3 and c5-depleted sera.

Okroj, Marcin LU ; Holmquist, Emelie LU ; King, Ben LU and Blom, Anna LU (2012) In PLoS ONE 7(10).
Abstract
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... (More)
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. (Less)
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author
organization
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Contribution to journal
publication status
published
subject
in
PLoS ONE
volume
7
issue
10
publisher
Public Library of Science
external identifiers
  • wos:000312385200095
  • pmid:23071769
  • scopus:84867391087
ISSN
1932-6203
DOI
10.1371/journal.pone.0047245
language
English
LU publication?
yes
id
2a974fd3-f08b-4b63-bb04-369a0763aae8 (old id 3160665)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/23071769?dopt=Abstract
date added to LUP
2012-11-01 13:45:11
date last changed
2017-01-01 06:16:27
@article{2a974fd3-f08b-4b63-bb04-369a0763aae8,
  abstract     = {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.},
  articleno    = {e47245},
  author       = {Okroj, Marcin and Holmquist, Emelie and King, Ben and Blom, Anna},
  issn         = {1932-6203},
  language     = {eng},
  number       = {10},
  publisher    = {Public Library of Science},
  series       = {PLoS ONE},
  title        = {Functional analyses of complement convertases using c3 and c5-depleted sera.},
  url          = {http://dx.doi.org/10.1371/journal.pone.0047245},
  volume       = {7},
  year         = {2012},
}