PMID- 33633190
OWN - NLM
STAT- MEDLINE
DCOM- 20211208
LR  - 20240331
IS  - 2045-2322 (Electronic)
IS  - 2045-2322 (Linking)
VI  - 11
IP  - 1
DP  - 2021 Feb 25
TI  - Specific domain V reduction of beta-2-glycoprotein I induces protein flexibility 
      and alters pathogenic antibody binding.
PG  - 4542
LID - 10.1038/s41598-021-84021-2 [doi]
LID - 4542
AB  - Beta-2-glycoprotein I (beta2GPI) is a blood protein and the major antigen in the 
      autoimmune disorder antiphospholipid syndrome (APS). beta2GPI exists mainly in 
      closed or open conformations and comprises of 11 disulfides distributed across 
      five domains. The terminal Cys288/Cys326 disulfide bond at domain V has been 
      associated with different cysteine redox states. The role of this disulfide bond 
      in conformational dynamics of this protein has not been investigated so far. 
      Here, we report on the enzymatic driven reduction by thioredoxin-1 (recycled by 
      Tris(2-carboxyethyl)phosphine; TCEP) of beta2GPI. Specific reduction was 
      demonstrated by Western blot and mass spectrometry analyses confirming majority 
      targeting to the fifth domain of beta2GPI. Atomic force microscopy images suggested 
      that reduced beta2GPI shows a slightly higher proportion of open conformation and is 
      more flexible compared to the untreated protein as confirmed by modelling 
      studies. We have determined a strong increase in the binding of pathogenic APS 
      autoantibodies to reduced beta2GPI as demonstrated by ELISA. Our study is relevant 
      for understanding the effect of beta2GPI reduction on the protein structure and its 
      implications for antibody binding in APS patients.
FAU - Buchholz, Ina
AU  - Buchholz I
AD  - Institute of Biochemistry, University of Greifswald, Greifswald, Germany.
AD  - ZIK HIKE, University of Greifswald, Greifswald, Germany.
FAU - McDonnell, Thomas
AU  - McDonnell T
AD  - Division of Biochemical Engineering, Bernard Katz Institute, University College 
      London, London, UK.
FAU - Nestler, Peter
AU  - Nestler P
AD  - Institute of Physics, University of Greifswald, Greifswald, Germany.
FAU - Tharad, Sudarat
AU  - Tharad S
AD  - Institute for Biophysics, University of Natural Resources and Life Sciences 
      Vienna, Vienna, Austria.
FAU - Kulke, Martin
AU  - Kulke M
AD  - Institute of Biochemistry, University of Greifswald, Greifswald, Germany.
FAU - Radziszewska, Anna
AU  - Radziszewska A
AD  - Centre for Adolescent Rheumatology Versus Arthritis at UCL, UCLH, GOSH, London, 
      UK.
AD  - Division of Medicine, Centre for Rheumatology, University College London, London, 
      UK.
FAU - Ripoll, Vera M
AU  - Ripoll VM
AD  - Division of Medicine, Centre for Rheumatology, University College London, London, 
      UK.
FAU - Schmidt, Frank
AU  - Schmidt F
AD  - Interfaculty Institute for Genetics and Functional Genomics, University of 
      Greifswald, Greifswald, Germany.
AD  - Proteomics Core, Weill Cornell Medicine-Qatar, Doha, Qatar.
FAU - Hammer, Elke
AU  - Hammer E
AD  - Interfaculty Institute for Genetics and Functional Genomics, University of 
      Greifswald, Greifswald, Germany.
AD  - DZHK (German Centre for Cardiovascular Research), Partner Site Greifswald, 
      Greifswald, Germany.
FAU - Toca-Herrera, Jose L
AU  - Toca-Herrera JL
AD  - Institute for Biophysics, University of Natural Resources and Life Sciences 
      Vienna, Vienna, Austria.
FAU - Rahman, Anisur
AU  - Rahman A
AD  - Division of Medicine, Centre for Rheumatology, University College London, London, 
      UK. anisur.rahman@ucl.ac.uk.
FAU - Delcea, Mihaela
AU  - Delcea M
AD  - Institute of Biochemistry, University of Greifswald, Greifswald, Germany. 
      delceam@uni-greifswald.de.
AD  - ZIK HIKE, University of Greifswald, Greifswald, Germany. 
      delceam@uni-greifswald.de.
AD  - DZHK (German Centre for Cardiovascular Research), Partner Site Greifswald, 
      Greifswald, Germany. delceam@uni-greifswald.de.
LA  - eng
GR  - MR/P017371/1/MRC_/Medical Research Council/United Kingdom
GR  - MRF_MRF-057-0004-RG-MCDO-C0800/MRF/MRF/United Kingdom
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20210225
PL  - England
TA  - Sci Rep
JT  - Scientific reports
JID - 101563288
RN  - 0 (Autoantibodies)
RN  - 0 (Disulfides)
RN  - 0 (Immunoglobulin G)
RN  - 0 (beta 2-Glycoprotein I)
RN  - K848JZ4886 (Cysteine)
SB  - IM
MH  - Autoantibodies/*chemistry/immunology
MH  - Cysteine/chemistry/metabolism
MH  - Disulfides/chemistry
MH  - Immunoglobulin G/chemistry/immunology
MH  - Microscopy, Atomic Force
MH  - Models, Molecular
MH  - Protein Binding/immunology
MH  - *Protein Conformation
MH  - *Protein Folding
MH  - *Protein Interaction Domains and Motifs
MH  - Structure-Activity Relationship
MH  - beta 2-Glycoprotein I/*chemistry/immunology/metabolism
PMC - PMC7907366
COIS- The authors declare no competing interests.
EDAT- 2021/02/27 06:00
MHDA- 2021/12/15 06:00
PMCR- 2021/02/25
CRDT- 2021/02/26 05:57
PHST- 2020/10/10 00:00 [received]
PHST- 2021/02/10 00:00 [accepted]
PHST- 2021/02/26 05:57 [entrez]
PHST- 2021/02/27 06:00 [pubmed]
PHST- 2021/12/15 06:00 [medline]
PHST- 2021/02/25 00:00 [pmc-release]
AID - 10.1038/s41598-021-84021-2 [pii]
AID - 84021 [pii]
AID - 10.1038/s41598-021-84021-2 [doi]
PST - epublish
SO  - Sci Rep. 2021 Feb 25;11(1):4542. doi: 10.1038/s41598-021-84021-2.