3-Phosphonopropionic Acids Inhibit Carboxypeptidase A As Multisubstrate Analogues or Transition-state Analogues

Overview

Journal Biochem J

Specialty Biochemistry

Date 1985 Nov 15

PMID 4084224

Citations 2

Authors

D Grobelny

U B Goli

R E Galardy

Affiliations

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Abstract

A series of phosphonic acid analogues of 2-benzylsuccinate were tested as inhibitors of carboxypeptidase A. The most potent of these, (2RS)-2-benzyl-3-phosphonopropionic acid, had a Ki of 0.22 +/- 0.05 microM, equipotent to (2RS)-2-benzylsuccinate and thus one of the most potent reversible inhibitors known for this enzyme. Lengthening by one methylene group to (2RS)-2-benzyl-4-phosphonobutyric acid increased the Ki to 370 +/- 60 microM. The monoethyl ester (2RS)-2-benzyl-3-(O-ethylphosphono)propionic acid was nearly as potent as (2RS)-2-benzyl-3-phosphonopropionic acid, with a Ki of 0.72 +/- 0.3 microM. The sulphur analogue of the monoethyl ester, 2-ambo-P-ambo-2-benzyl-3-(O-ethylthiophosphono)propionic acid, had a Ki of 2.1 +/- 0.6 microM, nearly as active as (2RS)-2-benzyl-3-(O-ethylphosphono)propionic acid. These phosphonic acids and esters could be considered to be multisubstrate inhibitors of carboxypeptidase A by virtue of their structural analogy with 2-benzylsuccinate. Alternatively, the tetrahedral hybridization at the phosphorus atom suggests that they could be mimicking a tetrahedral transition state for the enzyme-catalysed hydrolysis of substrate.

Citing Articles

Binding to thermolysin of phenolate-containing inhibitors necessitates a revised mechanism of catalysis.

Mock W, Aksamawati M Biochem J. 1994; 302 ( Pt 1):57-68.

PMID: 8068024 PMC: 1137190. DOI: 10.1042/bj3020057.

Ionization states of the complex formed between 2-benzyl-3-phosphonopropionic acid and carboxypeptidase A.

Goli U, Grobelny D, Galardy R Biochem J. 1988; 254(3):847-53.

PMID: 3196297 PMC: 1135160. DOI: 10.1042/bj2540847.

References

Schechter I, Berger A . On the active site of proteases. 3. Mapping the active site of papain; specific peptide inhibitors of papain. Biochem Biophys Res Commun. 1968; 32(5):898-902. DOI: 10.1016/0006-291x(68)90326-4. View

Bandurski R, AXELROD B . The chromatographic identification of some biologically important phosphate esters. J Biol Chem. 1951; 193(1):405-10. View

Byers L, Wolfenden R . Binding of the by-product analog benzylsuccinic acid by carboxypeptidase A. Biochemistry. 1973; 12(11):2070-8. DOI: 10.1021/bi00735a008. View

Suh J, Kaiser E . pH dependence of the nitrotyrosine-248 and arsanilazotyrosine-248 carboxypeptidase A catalyzed hydrolysis of O-(trans-p-chlorocinnamoyl)-L-beta-phenyllactate. J Am Chem Soc. 1976; 98(7):1940-7. DOI: 10.1021/ja00423a048. View

Weaver L, Kester W, Matthews B . A crystallographic study of the complex of phosphoramidon with thermolysin. A model for the presumed catalytic transition state and for the binding of extended substances. J Mol Biol. 1977; 114(1):119-32. DOI: 10.1016/0022-2836(77)90286-8. View

Bolognesi M, Matthews B . Binding of the biproduct analog L-benzylsuccinic acid to thermolysin determined by X-ray crystallography. J Biol Chem. 1979; 254(3):634-9. View

ONDETTI M, CONDON M, Reid J, SABO E, Cheung H, Cushman D . Design of potent and specific inhibitors of carboxypeptidases A and B. Biochemistry. 1979; 18(8):1427-30. DOI: 10.1021/bi00575a006. View

Kam C, Nishino N, POWERS J . Inhibition of thermolysin and carboxypeptidase A by phosphoramidates. Biochemistry. 1979; 18(14):3032-8. DOI: 10.1021/bi00581a019. View

Nishino N, POWERS J . Design of potent reversible inhibitors for thermolysin. Peptides containing zinc coordinating ligands and their use in affinity chromatography. Biochemistry. 1979; 18(20):4340-7. DOI: 10.1021/bi00587a012. View

10.

Holmquist B, Vallee B . Metal-coordinating substrate analogs as inhibitors of metalloenzymes. Proc Natl Acad Sci U S A. 1979; 76(12):6216-20. PMC: 411834. DOI: 10.1073/pnas.76.12.6216. View

11.

Monzingo A, Matthews B . Structure of a mercaptan-thermolysin complex illustrates mode of inhibition of zinc proteases by substrate-analogue mercaptans. Biochemistry. 1982; 21(14):3390-4. DOI: 10.1021/bi00257a022. View

12.

Galardy R . Inhibition of angiotensin converting enzyme by phosphoramidates and polyphosphates. Biochemistry. 1982; 21(23):5777-81. DOI: 10.1021/bi00266a008. View

13.

Galardy R, Grobelny D . Inhibition of collagenase from Clostridium histolyticum by phosphoric and phosphonic amides. Biochemistry. 1983; 22(19):4556-61. DOI: 10.1021/bi00288a032. View

14.

Bartlett P, Marlowe C . Phosphonamidates as transition-state analogue inhibitors of thermolysin. Biochemistry. 1983; 22(20):4618-24. DOI: 10.1021/bi00289a002. View

15.

Galardy R, Kortylewicz Z . Inhibition of carboxypeptidase A by aldehyde and ketone substrate analogues. Biochemistry. 1984; 23(9):2083-7. DOI: 10.1021/bi00304a032. View

16.

Poncz L, Gerken T, Dearborn D, Grobelny D, Galardy R . Inhibition of the elastase of Pseudomonas aeruginosa by N alpha-phosphoryl dipeptides and kinetics of spontaneous hydrolysis of the inhibitors. Biochemistry. 1984; 23(12):2766-72. DOI: 10.1021/bi00307a036. View

17.

Galardy R, Kortylewicz Z . Inhibitors of angiotensin-converting enzyme containing a tetrahedral arsenic atom. Biochem J. 1985; 226(2):447-54. PMC: 1144731. DOI: 10.1042/bj2260447. View

18.

Quiocho F, Lipscomb W . Carboxypeptidase A: a protein and an enzyme. Adv Protein Chem. 1971; 25:1-78. DOI: 10.1016/s0065-3233(08)60278-8. View