PMID- 32946534
OWN - NLM
STAT- MEDLINE
DCOM- 20201105
LR  - 20210212
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 15
IP  - 9
DP  - 2020
TI  - Binding of omeprazole to protein targets identified by monoclonal antibodies.
PG  - e0239464
LID - 10.1371/journal.pone.0239464 [doi]
LID - e0239464
AB  - Omeprazole is the most commonly used proton pump inhibitor (PPI), a class of 
      medications whose therapeutic mechanism of action involves formation of a 
      disulfide linkage to cysteine residues in the H+/K+ ATPase pump on gastric 
      secretory cells. Covalent linkage between the sole sulfur group of omeprazole and 
      selected cysteine residues of the pump protein results in inhibition of acid 
      secretion in the stomach, an effect that ameliorates gastroesophageal reflux and 
      peptic ulcer disease. PPIs, though useful for specific conditions when used 
      transiently, are associated with diverse untoward effects when used long term. 
      The mechanisms underlying these potential off-target effects remain unclear. PPIs 
      may, in fact, interact with non-canonical target proteins (non-pump molecules) 
      resulting in unexpected pathophysiological effects, but few studies describe 
      off-target PPI binding. Here, we describe successful cloning of monoclonal 
      antibodies against protein-bound omeprazole. We developed and used monoclonal 
      antibodies to characterize the protein target range of omeprazole, stability of 
      omeprazole-bound proteins, and the involvement of cysteines in binding of 
      omeprazole to targets. We demonstrate that a wide range of diverse proteins are 
      targeted by omeprazole. Protein complexes, detected by Western blotting, are 
      resistant to heat, detergents, and reducing agents. Reaction of omeprazole occurs 
      with cysteine-free proteins, is not fully inhibited by cysteine alkylation, 
      occurs at neutral pH, and induces protein multimerization. At least two other 
      clinically used PPIs, rabeprazole and tenatoprazole, are capable of binding to 
      proteins in a similar fashion. We conclude that omeprazole binds to multiple 
      proteins and is capable of forming highly stable complexes that are not dependent 
      on disulfide linkages between the drug and protein targets. Further studies made 
      possible by these antibodies may shed light on whether PPI-protein complexes 
      underlie off-target untoward effects of chronic PPI use.
FAU - Cartee, Naw May Pearl
AU  - Cartee NMP
AD  - Department of Neurology, University of Michigan, Ann Arbor, MI, United States of 
      America.
AD  - Department of Veterans Affairs, Neurology Service, VA Ann Arbor Healthcare 
      System, Ann Arbor, MI, United States of America.
FAU - Wang, Michael M
AU  - Wang MM
AUID- ORCID: 0000-0002-5670-2496
AD  - Department of Neurology, University of Michigan, Ann Arbor, MI, United States of 
      America.
AD  - Department of Veterans Affairs, Neurology Service, VA Ann Arbor Healthcare 
      System, Ann Arbor, MI, United States of America.
AD  - Molecular and Integrative Physiology, University of Michigan, Ann Arbor, MI, 
      United States of America.
LA  - eng
GR  - I01 BX003824/BX/BLRD VA/United States
GR  - I01 BX003855/BX/BLRD VA/United States
GR  - R01 NS099160/NS/NINDS NIH HHS/United States
GR  - R21 NS099783/NS/NINDS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20200918
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Antibodies, Monoclonal, Murine-Derived)
RN  - 0 (Multiprotein Complexes)
RN  - 0 (Proton Pump Inhibitors)
RN  - K848JZ4886 (Cysteine)
RN  - KG60484QX9 (Omeprazole)
SB  - IM
MH  - Animals
MH  - Antibodies, Monoclonal, Murine-Derived/*metabolism
MH  - Binding Sites
MH  - Cysteine/chemistry
MH  - HEK293 Cells
MH  - Humans
MH  - Mice
MH  - Mice, Inbred BALB C
MH  - Mice, Inbred C57BL
MH  - Multiprotein Complexes/chemistry/metabolism
MH  - Omeprazole/chemistry/*immunology/*metabolism
MH  - Protein Binding
MH  - Proton Pump Inhibitors/chemistry/*immunology/*metabolism
PMC - PMC7500594
COIS- The authors have declared that no competing interests exist.
EDAT- 2020/09/19 06:00
MHDA- 2020/11/06 06:00
PMCR- 2020/09/18
CRDT- 2020/09/18 17:12
PHST- 2020/05/07 00:00 [received]
PHST- 2020/09/08 00:00 [accepted]
PHST- 2020/09/18 17:12 [entrez]
PHST- 2020/09/19 06:00 [pubmed]
PHST- 2020/11/06 06:00 [medline]
PHST- 2020/09/18 00:00 [pmc-release]
AID - PONE-D-20-13524 [pii]
AID - 10.1371/journal.pone.0239464 [doi]
PST - epublish
SO  - PLoS One. 2020 Sep 18;15(9):e0239464. doi: 10.1371/journal.pone.0239464. 
      eCollection 2020.