Lansoprazole inhibits the cysteine protease legumain by binding to the active site

Tatjana Bosnjak, Rigmor Solberg, Paya Diana Hemati, Abbas Jafari, Moustapha Kassem, Harald Thidemann Johansen*

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Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

Proton pump inhibitors (PPIs) are prodrugs used in the treatment of peptic ulcer diseases. Once activated by acidic pH, the PPIs subsequently inhibit the secretion of gastric acid by covalently forming disulphide bonds with the SH groups of the parietal proton pump, that is the H + /K + -ATPase. Long-term use of PPIs has been associated with numerous adverse effects, including bone fractures. Considering the mechanism of activation, PPIs could also be active in acidic micro-environments such as in lysosomes, tumours and bone resorption sites. We suggested that the SH group in the active site of cysteine proteases could be susceptible for inhibition by PPIs. In this study, the inhibition by lansoprazole was shown on the cysteine proteases legumain and cathepsin B by incubating purified proteases or cell lysates with lansoprazole at different concentrations and pH conditions. The mechanism of legumain inhibition was shown to be a direct interaction of lansoprazole with the SH group in the active site, and thus blocking binding of the legumain-selective activity-based probe MP-L01. Lansoprazole was also shown to inhibit both legumain and cathepsin B in various cell models like HEK293, monoclonal legumain over-expressing HEK293 cells (M38L) and RAW264.7 macrophages, but not in human bone marrow-derived skeletal (mesenchymal) stem cells (hBMSC-TERT). During hBMSC-TERT differentiation to osteoblasts, lansoprazole inhibited legumain secretion, alkaline phosphatase activity, but had no effects on in vitro mineralization capacity. In conclusion, lansoprazole acts as a direct covalent inhibitor of cysteine proteases via disulphide bonds with the SH group in the protease active site. Such inhibition of cysteine proteases could explain some of the off-target effects of PPIs.

OriginalsprogEngelsk
TidsskriftBasic & Clinical Pharmacology & Toxicology
Vol/bind125
Udgave nummer2
Sider (fra-til)89-99
ISSN1742-7835
DOI
StatusUdgivet - aug. 2019

Fingeraftryk

asparaginylendopeptidase
Lansoprazole
Cysteine Proteases
Proton Pump Inhibitors
Catalytic Domain
Cathepsin B
Bone
Disulfides
Peptide Hydrolases
Cysteine Proteinase Inhibitors
Proton Pumps
Macrophages
HEK293 Cells
Gastric Acid
Osteoblasts
Bone Fractures
Prodrugs
Stem cells
Mesenchymal Stromal Cells
Alkaline Phosphatase

Citer dette

Bosnjak, Tatjana ; Solberg, Rigmor ; Hemati, Paya Diana ; Jafari, Abbas ; Kassem, Moustapha ; Johansen, Harald Thidemann. / Lansoprazole inhibits the cysteine protease legumain by binding to the active site. I: Basic & Clinical Pharmacology & Toxicology. 2019 ; Bind 125, Nr. 2. s. 89-99.
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title = "Lansoprazole inhibits the cysteine protease legumain by binding to the active site",
abstract = "Proton pump inhibitors (PPIs) are prodrugs used in the treatment of peptic ulcer diseases. Once activated by acidic pH, the PPIs subsequently inhibit the secretion of gastric acid by covalently forming disulphide bonds with the SH groups of the parietal proton pump, that is the H + /K + -ATPase. Long-term use of PPIs has been associated with numerous adverse effects, including bone fractures. Considering the mechanism of activation, PPIs could also be active in acidic micro-environments such as in lysosomes, tumours and bone resorption sites. We suggested that the SH group in the active site of cysteine proteases could be susceptible for inhibition by PPIs. In this study, the inhibition by lansoprazole was shown on the cysteine proteases legumain and cathepsin B by incubating purified proteases or cell lysates with lansoprazole at different concentrations and pH conditions. The mechanism of legumain inhibition was shown to be a direct interaction of lansoprazole with the SH group in the active site, and thus blocking binding of the legumain-selective activity-based probe MP-L01. Lansoprazole was also shown to inhibit both legumain and cathepsin B in various cell models like HEK293, monoclonal legumain over-expressing HEK293 cells (M38L) and RAW264.7 macrophages, but not in human bone marrow-derived skeletal (mesenchymal) stem cells (hBMSC-TERT). During hBMSC-TERT differentiation to osteoblasts, lansoprazole inhibited legumain secretion, alkaline phosphatase activity, but had no effects on in vitro mineralization capacity. In conclusion, lansoprazole acts as a direct covalent inhibitor of cysteine proteases via disulphide bonds with the SH group in the protease active site. Such inhibition of cysteine proteases could explain some of the off-target effects of PPIs.",
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author = "Tatjana Bosnjak and Rigmor Solberg and Hemati, {Paya Diana} and Abbas Jafari and Moustapha Kassem and Johansen, {Harald Thidemann}",
year = "2019",
month = "8",
doi = "10.1111/bcpt.13230",
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pages = "89--99",
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Lansoprazole inhibits the cysteine protease legumain by binding to the active site. / Bosnjak, Tatjana; Solberg, Rigmor; Hemati, Paya Diana; Jafari, Abbas; Kassem, Moustapha; Johansen, Harald Thidemann.

I: Basic & Clinical Pharmacology & Toxicology, Bind 125, Nr. 2, 08.2019, s. 89-99.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Lansoprazole inhibits the cysteine protease legumain by binding to the active site

AU - Bosnjak, Tatjana

AU - Solberg, Rigmor

AU - Hemati, Paya Diana

AU - Jafari, Abbas

AU - Kassem, Moustapha

AU - Johansen, Harald Thidemann

PY - 2019/8

Y1 - 2019/8

N2 - Proton pump inhibitors (PPIs) are prodrugs used in the treatment of peptic ulcer diseases. Once activated by acidic pH, the PPIs subsequently inhibit the secretion of gastric acid by covalently forming disulphide bonds with the SH groups of the parietal proton pump, that is the H + /K + -ATPase. Long-term use of PPIs has been associated with numerous adverse effects, including bone fractures. Considering the mechanism of activation, PPIs could also be active in acidic micro-environments such as in lysosomes, tumours and bone resorption sites. We suggested that the SH group in the active site of cysteine proteases could be susceptible for inhibition by PPIs. In this study, the inhibition by lansoprazole was shown on the cysteine proteases legumain and cathepsin B by incubating purified proteases or cell lysates with lansoprazole at different concentrations and pH conditions. The mechanism of legumain inhibition was shown to be a direct interaction of lansoprazole with the SH group in the active site, and thus blocking binding of the legumain-selective activity-based probe MP-L01. Lansoprazole was also shown to inhibit both legumain and cathepsin B in various cell models like HEK293, monoclonal legumain over-expressing HEK293 cells (M38L) and RAW264.7 macrophages, but not in human bone marrow-derived skeletal (mesenchymal) stem cells (hBMSC-TERT). During hBMSC-TERT differentiation to osteoblasts, lansoprazole inhibited legumain secretion, alkaline phosphatase activity, but had no effects on in vitro mineralization capacity. In conclusion, lansoprazole acts as a direct covalent inhibitor of cysteine proteases via disulphide bonds with the SH group in the protease active site. Such inhibition of cysteine proteases could explain some of the off-target effects of PPIs.

AB - Proton pump inhibitors (PPIs) are prodrugs used in the treatment of peptic ulcer diseases. Once activated by acidic pH, the PPIs subsequently inhibit the secretion of gastric acid by covalently forming disulphide bonds with the SH groups of the parietal proton pump, that is the H + /K + -ATPase. Long-term use of PPIs has been associated with numerous adverse effects, including bone fractures. Considering the mechanism of activation, PPIs could also be active in acidic micro-environments such as in lysosomes, tumours and bone resorption sites. We suggested that the SH group in the active site of cysteine proteases could be susceptible for inhibition by PPIs. In this study, the inhibition by lansoprazole was shown on the cysteine proteases legumain and cathepsin B by incubating purified proteases or cell lysates with lansoprazole at different concentrations and pH conditions. The mechanism of legumain inhibition was shown to be a direct interaction of lansoprazole with the SH group in the active site, and thus blocking binding of the legumain-selective activity-based probe MP-L01. Lansoprazole was also shown to inhibit both legumain and cathepsin B in various cell models like HEK293, monoclonal legumain over-expressing HEK293 cells (M38L) and RAW264.7 macrophages, but not in human bone marrow-derived skeletal (mesenchymal) stem cells (hBMSC-TERT). During hBMSC-TERT differentiation to osteoblasts, lansoprazole inhibited legumain secretion, alkaline phosphatase activity, but had no effects on in vitro mineralization capacity. In conclusion, lansoprazole acts as a direct covalent inhibitor of cysteine proteases via disulphide bonds with the SH group in the protease active site. Such inhibition of cysteine proteases could explain some of the off-target effects of PPIs.

KW - asparaginyl endopeptidase

KW - cathepsin B

KW - lansoprazole

KW - legumain

KW - proton pump inhibitor

U2 - 10.1111/bcpt.13230

DO - 10.1111/bcpt.13230

M3 - Journal article

C2 - 30916878

AN - SCOPUS:85064481633

VL - 125

SP - 89

EP - 99

JO - Basic & Clinical Pharmacology & Toxicology

JF - Basic & Clinical Pharmacology & Toxicology

SN - 1742-7835

IS - 2

ER -