Molecular Interactions and Implications of Aldose Reductase Inhibition by PGA1 and Clinically Used Prostaglandins.
Aldose reductase (AKR1B1) is a critical drug target because of its involvement in diabetic complications, inflammation, and tumorigenesis. However, to date, development of clinically useful inhibitors has been largely unsuccessful. Cyclopentenone prostaglandins (cyPGs) are reactive lipid mediators that bind covalently to proteins and exert anti-inflammatory and antiproliferative effects in numerous settings. By pursuing targets for modification by cyPGs we have found that the cyPG PGA1 binds to and inactivates AKR1B1. A PGA1-AKR1B1 adduct was observed, both by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and by SDS-PAGE using biotinylated PGA1 (PGA1-B). Insight into the molecular interactions between AKR1B1 and PGA1 was advanced by molecular modeling. This anticipated the addition of PGA1 to active site Cys298 and the potential reversibility of the adduct, which was supported experimentally. Indeed, loss of biotin label from the AKR1B1-PGA1-B adduct was favored by glutathione, indicating a retro-Michael reaction, which unveils new implications of cyPG-protein interaction. PGA1 elicited only marginal inhibition of aldehyde reductase (AKR1A1), considered responsible for the severe adverse effects of many AKR1B1 inhibitors. Interestingly, other prostaglandins (PGs) inhibited the enzyme, including non-electrophilic PGE1 and PGE2, currently used in clinical practice. Moreover, both PGA1 and PGE1 reduced the formation of sorbitol in an ex-vivo model of diabetic cataract to an extent comparable to that attained by the known AKR inhibitor epalrestat. Taken together, these results highlight the role of PGs as AKR1B1 inhibitors and the interest in PG-related molecules as leads for the development of novel pharmacological tools.
Chemical and Physical Biology Department, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain (B.D.-D., F.J.S.-G., T.Z., D.P.-S.); Department of Biomedical Sciences, Universidad de Alcalá, Madrid, Spain (P.A.S.-M., F.G.); Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, Bratislava, Slovakia (I.M., J.B., M.S.); Department of Pharmacology, University of Extremadura, Cáceres, Spain (E.G.-M., J.A.G.A.); Centro Nacional de Biotecnología, Madrid, Spain (S.G.).
Beatriz
B
Díez-Dacal
Chemical and Physical Biology Department, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain (B.D.-D., F.J.S.-G., T.Z., D.P.-S.); Department of Biomedical Sciences, Universidad de Alcalá, Madrid, Spain (P.A.S.-M., F.G.); Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, Bratislava, Slovakia (I.M., J.B., M.S.); Department of Pharmacology, University of Extremadura, Cáceres, Spain (E.G.-M., J.A.G.A.); Centro Nacional de Biotecnología, Madrid, Spain (S.G.).
Francisco J
FJ
Sánchez-Gómez
Chemical and Physical Biology Department, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain (B.D.-D., F.J.S.-G., T.Z., D.P.-S.); Department of Biomedical Sciences, Universidad de Alcalá, Madrid, Spain (P.A.S.-M., F.G.); Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, Bratislava, Slovakia (I.M., J.B., M.S.); Department of Pharmacology, University of Extremadura, Cáceres, Spain (E.G.-M., J.A.G.A.); Centro Nacional de Biotecnología, Madrid, Spain (S.G.).
Pedro A
PA
Sánchez-Murcia
Chemical and Physical Biology Department, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain (B.D.-D., F.J.S.-G., T.Z., D.P.-S.); Department of Biomedical Sciences, Universidad de Alcalá, Madrid, Spain (P.A.S.-M., F.G.); Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, Bratislava, Slovakia (I.M., J.B., M.S.); Department of Pharmacology, University of Extremadura, Cáceres, Spain (E.G.-M., J.A.G.A.); Centro Nacional de Biotecnología, Madrid, Spain (S.G.).
Ivana
I
Milackova
Chemical and Physical Biology Department, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain (B.D.-D., F.J.S.-G., T.Z., D.P.-S.); Department of Biomedical Sciences, Universidad de Alcalá, Madrid, Spain (P.A.S.-M., F.G.); Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, Bratislava, Slovakia (I.M., J.B., M.S.); Department of Pharmacology, University of Extremadura, Cáceres, Spain (E.G.-M., J.A.G.A.); Centro Nacional de Biotecnología, Madrid, Spain (S.G.).
Tahl
T
Zimmerman
Chemical and Physical Biology Department, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain (B.D.-D., F.J.S.-G., T.Z., D.P.-S.); Department of Biomedical Sciences, Universidad de Alcalá, Madrid, Spain (P.A.S.-M., F.G.); Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, Bratislava, Slovakia (I.M., J.B., M.S.); Department of Pharmacology, University of Extremadura, Cáceres, Spain (E.G.-M., J.A.G.A.); Centro Nacional de Biotecnología, Madrid, Spain (S.G.).
Jana
J
Ballekova
Chemical and Physical Biology Department, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain (B.D.-D., F.J.S.-G., T.Z., D.P.-S.); Department of Biomedical Sciences, Universidad de Alcalá, Madrid, Spain (P.A.S.-M., F.G.); Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, Bratislava, Slovakia (I.M., J.B., M.S.); Department of Pharmacology, University of Extremadura, Cáceres, Spain (E.G.-M., J.A.G.A.); Centro Nacional de Biotecnología, Madrid, Spain (S.G.).
Elena
E
García-Martín
Chemical and Physical Biology Department, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain (B.D.-D., F.J.S.-G., T.Z., D.P.-S.); Department of Biomedical Sciences, Universidad de Alcalá, Madrid, Spain (P.A.S.-M., F.G.); Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, Bratislava, Slovakia (I.M., J.B., M.S.); Department of Pharmacology, University of Extremadura, Cáceres, Spain (E.G.-M., J.A.G.A.); Centro Nacional de Biotecnología, Madrid, Spain (S.G.).
José A G
JA
Agúndez
Chemical and Physical Biology Department, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain (B.D.-D., F.J.S.-G., T.Z., D.P.-S.); Department of Biomedical Sciences, Universidad de Alcalá, Madrid, Spain (P.A.S.-M., F.G.); Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, Bratislava, Slovakia (I.M., J.B., M.S.); Department of Pharmacology, University of Extremadura, Cáceres, Spain (E.G.-M., J.A.G.A.); Centro Nacional de Biotecnología, Madrid, Spain (S.G.).
Severine
S
Gharbi
Chemical and Physical Biology Department, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain (B.D.-D., F.J.S.-G., T.Z., D.P.-S.); Department of Biomedical Sciences, Universidad de Alcalá, Madrid, Spain (P.A.S.-M., F.G.); Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, Bratislava, Slovakia (I.M., J.B., M.S.); Department of Pharmacology, University of Extremadura, Cáceres, Spain (E.G.-M., J.A.G.A.); Centro Nacional de Biotecnología, Madrid, Spain (S.G.).
Federico
F
Gago
Chemical and Physical Biology Department, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain (B.D.-D., F.J.S.-G., T.Z., D.P.-S.); Department of Biomedical Sciences, Universidad de Alcalá, Madrid, Spain (P.A.S.-M., F.G.); Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, Bratislava, Slovakia (I.M., J.B., M.S.); Department of Pharmacology, University of Extremadura, Cáceres, Spain (E.G.-M., J.A.G.A.); Centro Nacional de Biotecnología, Madrid, Spain (S.G.).
Milan
M
Stefek
Chemical and Physical Biology Department, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain (B.D.-D., F.J.S.-G., T.Z., D.P.-S.); Department of Biomedical Sciences, Universidad de Alcalá, Madrid, Spain (P.A.S.-M., F.G.); Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, Bratislava, Slovakia (I.M., J.B., M.S.); Department of Pharmacology, University of Extremadura, Cáceres, Spain (E.G.-M., J.A.G.A.); Centro Nacional de Biotecnología, Madrid, Spain (S.G.) dperezsala@cib.csic.es.
Dolores
http://orcid.org/0000-0003-0600-665X
D
Pérez-Sala
D000449:Aldehyde Reductase / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D000818:Animals; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D008297:Male; D011453:Prostaglandins / Q000378:metabolism / Q000494:pharmacology; D011454:Prostaglandins A / Q000378:metabolism* / Q000494:pharmacology*; D011485:Protein Binding / Q000502:physiology; D051381:Rats; D017208:Rats, Wistar
D004791:Enzyme Inhibitors; D011453:Prostaglandins; D011454:Prostaglandins A; C576693:Akr1b1 protein, rat; D000449:Aldehyde Reductase; C100573:prostaglandin A1