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Analysis of the interaction between human steroid 21-hydroxylase and various monoclonal antibodies using comparative structural modelling
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Miguel, R. N., Chen, S., Nikfarjam, L., Kominami, S., Carpenter, Byron, Dal Pra, C., Betterle, C., Zanchetta, R., Nakamatsu , T., Powell, M., Hewer, R., Blundell , T. L., Smith, B. R. and Furmaniak, J. (2005) Analysis of the interaction between human steroid 21-hydroxylase and various monoclonal antibodies using comparative structural modelling. European Journal of Endocrinology, 153 (6). pp. 949-961. doi:10.1530/eje.1.02044 ISSN 0804-4643.
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Official URL: http://dx.doi.org/10.1530/eje.1.02044
Abstract
Objective: To study the interaction between human steroid 21-hydroxylase (21-OH) and monoclonal antibodies (MAbs) to 21-OH directed to 3 different epitopes recognised by 21-OH autoantibodies characteristic of autoimmune Addison’s disease.
Design: Build comparative structural models of 21-OH, 21-OH MAbs and complexes of 21-OH–21-OH MAbs and study the effects of 21-OH MAbs on 21-OH enzyme activity. Then, analyse the relationship between sites important for binding of 21-OH MAbs and 21-OH autoantibodies and sites important for 21-OH enzyme activity.
Methods: Variable (V) regions of 21-OH MAbs (M21-OH1, M21-OH3, M21-OH5) were sequenced and models of the MAbs built using structures of antibodies in the database as templates. A comparative model of 21-OH was built using the crystal structure of rabbit cytochrome p450 2c5/3LVdH as template. 21-OH enzyme activity was measured in terms of conversion of [3H]progesterone to deoxycorticosterone and the effect of purified MAb IgGs on 21-OH enzyme activity was assessed.
Results: M21-OH1, M21-OH3 and control MAb had no effect on 21-OH enzyme activity with 88.8% ± 24% (n = 6), 86.7% ± 7.6% (n = 6) and 86.5% ± 10.6% (n = 6) of activity remaining in the presence of the respective IgGs. This was consistent with the epitopes for M21-OH1 and M21-OH3 being located distant from 21-OH enzyme active sites in our 21-OH model. The epitope for M21-OH5 which inhibited 21-OH enzyme activity (48.5 ± 8.3% activity remaining; P < 0.001 compared with control MAb IgG) was found close to the redox protein binding site in our 21-OH model.
Conclusions: A comparative model of 21-OH has been produced. Analysis of experimental data in the context of the model suggests that M21-OH5 inhibits 21-OH enzyme activity through interference with redox protein binding.
Item Type: | Journal Article | ||||||
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | ||||||
Journal or Publication Title: | European Journal of Endocrinology | ||||||
Publisher: | BioScientifica Ltd. | ||||||
ISSN: | 0804-4643 | ||||||
Official Date: | 2005 | ||||||
Dates: |
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Volume: | 153 | ||||||
Number: | 6 | ||||||
Page Range: | pp. 949-961 | ||||||
DOI: | 10.1530/eje.1.02044 | ||||||
Status: | Peer Reviewed | ||||||
Publication Status: | Published | ||||||
Access rights to Published version: | Restricted or Subscription Access |
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