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Esterase-Like Activity of Serum Albumin: Characterization of Its Structural Chemistry Using p-Nitrophenyl Esters as Substrates

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Abstract

Purpose. To elucidate the catalytic mechanism of the esterase-like activity of serum albumin (SA), the reactivity of SA from six species was investigated using p-nitrophenyl esters as model substrates.

Methods. The effect of pH and the energetic and thermodynamic profiles of SA were determined for all species for p-nitrophenyl acetate (PNPA). Then, kinetic and thermodynamic studies using a series of p- and o-nitrophenyl esters with different side chains and human SA (HSA) were carried out. The influence of deuterium oxide was also evaluated. Finally, the information gained was used to construct a computer model of the structural chemistry of the reaction.

Results. The pH profiles suggest that the nucleophilic character of the catalytic residue (Tyr-411 in the case of HSA) is essential for activity. This k cat-dependent activity was found to increase with a decrease in the activation free energy change (ΔG). Hence, the magnitude of ΔG, which is dependent on activation entropy change (ΔS), as calculated from the thermodynamic analysis, can be regarded as an indicator of hydrolytic activity. It indicates that p-nitrophenyl propionate (PNPP) is the best substrate by evaluating the reactions of nitrophenyl esters with HSA. The findings here indicate that deuterium oxide has no significant effect on the rate of hydrolysis of PNPA by HSA.

Conclusions. The results are consistent with a scenario in which HSA becomes acylated due to a nucleophilic attack by Tyr-411 on the substrate and then is deacylated by general acid or base catalysis with the participation of water.

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references

  1. T. Peter Jr. All About Albumin, Biochemistry, Genetics, and Medical Applications, Academic Press, San Diego, 1996.

    Google Scholar 

  2. G. E. Means and M. L. Bender. Acetylation of human serum albumin by p-nitrophenyl acetate. Biochemistry 14:4989-4994 (1975).

    Google Scholar 

  3. M. A. Sogorb, A. Monroy, and E. Vilanova. Chicken serum albumin hydrolyzes dichlorophenyl phosphoramidates by a mechanism based on transient phosphorylation. Chem. Res. Toxicol. 11:1441-1446 (1998).

    Google Scholar 

  4. K. Ikeda and Y. Kurono. Enzymatic activity and drug binding activity of human serum albumin. Yakugaku Zasshi 106:841-855 (1986).

    Google Scholar 

  5. C. Y. Quon, K. Mai, G. Patil, and H. F. Stampfli. Species differences in the stereoselective hydrolysis of esmolol by blood esterases. Drug Metab. Dispos. 16:425-428 (1988).

    Google Scholar 

  6. T. Satoh, M. Hosokawa, R. Atsumi, W. Suzuki, H. Hakusui, and E. Nagai. Metabolic activation of CPT-11, 7-ethyl-10-4-(1-piperidino)-1-piperidino carbonyloxycamptothecin, a novel antitumor agent, by carboxylesterase. Biol. Pharm. Bull. 17:662-664 (1994).

    Google Scholar 

  7. A. Awad-Elkarim and G. E. Means. The reactivity of p-nitrophenyl acetate with serum albumin. Comp. Biochem. Physiol. 91B:267-272 (1988).

    Google Scholar 

  8. X. M. He and D. C. Carter. Atomic structure and chemistry of human serum albumin. Nature 358:209-215 (1992).

    Google Scholar 

  9. J. X. Ho, E. W. Holowachuk, E. J. Norton, P. D. Twigg, and D. C. Carter. X-ray and primary structure of horse serum albumin (Equus caballus) at 0.27-nm resolution. Eur. J. Biochem. 215:205-212 (1993).

    Google Scholar 

  10. H. Watanabe, S. Tanase, K. Nakajou, T. Maruyama, U. Kragh-Hansen, and M. Otagiri. Role of Arg-410 and Tyr-411 in human serum albumin for ligand binding and esterase-like activity. Biochem. J. 349:813-819 (2000).

    Google Scholar 

  11. R. F. Chen. Removal of fatty acids from serum albumin by charcoal treatment. J. Biol. Chem. 242:173-181 (1967).

    Google Scholar 

  12. Y. Kurono, T. Maki, T. Yotsuyanagi, and K. Ikeda. Esterase-like activity of human serum albumin: structure-activity relationships for the reactions with phenyl acetates and p-nitrophenyl esters. Chem. Pharm. Bull. 27:2781-2786 (1979).

    Google Scholar 

  13. N. Ohta, Y. Kurono, and K. Ikeda. Esterase-like activity of human serum albumin II: reaction with N-trans-cinnamoylimidazoles. J. Pharm. Sci. 72:385-388 (1983).

    Google Scholar 

  14. A. Fersht. Structure and Mechanism in Protein Science, Freeman, New York, 1998.

    Google Scholar 

  15. H. Tsujishita and S. Hirono. CAMDAS: an automated conformational analysis system using molecular dynamics. Conformational analyzer with molecular dynamics and sampling. J. Comput. Aided Mol. Design 11:305-315 (1997).

    Google Scholar 

  16. M. Clark, R. D. Cramer III, and N. Van Opdenbosch. Validation of the general purpose Tripos 5.2 force field. J. Comp. Chem. 10:982-1012 (1989).

    Google Scholar 

  17. S. Curry, H. Mandelkow, P. Brick, and N. Franks. Crystal structure of human serum albumin complexed with fatty acid reveals an asymmetric distribution of binding sites. Nature Struct. Biol. 5:827-835 (1998).

    Google Scholar 

  18. SYBYL 6.6.2, Tripos Inc., St. Louis, MO, 2000.

  19. S. J. Weiner, P. A. Kollman, D. T. Nguyen, and D. A. Case. An all atom force field for simulations of proteins and nucleic acids. J. Comp. Chem. 7:230-252 (1986).

    Google Scholar 

  20. J. Gasteiger and M. Marsili. Iterative partial equalization of orbital electronegativity—a rapid access to atomic charges. Tetrahedron 36:3219-3228 (1980).

    Google Scholar 

  21. M. Marsili and J. Gasteiger. Croat. Chem. Acta 53:601-614 (1980).

    Google Scholar 

  22. J. Gasteiger and M. Marsili. Prediction of proton magnetic resonance shifts: the dependence on hydrogen charges obtained by iterative partial equalization of orbital electronegativity. Organ. Magn. Reson. 15:353-360 (1981).

    Google Scholar 

  23. W. P. Purcell and J. A. Singer. A brief review and table of semiempirical parameters used in the Hüeckel molecular orbital method. J. Chem. Eng. Data 12:235-246 (1967).

    Google Scholar 

  24. UNITY 4.1.2, Tripos Inc., St. Louis, MO, 2000.

  25. G. P. Ford and B. Wang. New approach to the rapid semiempirical calculation of molecular electrostatic potentials based on the AM1 wave function: Comparison with Ab Initio HF/6-31G* results. J. Comp. Chem. 14:1101-1111 (1993).

    Google Scholar 

  26. B. Wang and G. P. Ford. Atomic charges derived from a fast and accurate method for electrostatic potentials based on modified AM1 calculations. J. Comp. Chem. 15:200-207 (1994).

    Google Scholar 

  27. CS ChemOffice Ultra 2000 Enhanced 5.5, CambridgeSoft, Cambridge, MA, 2000.

  28. M. Dockal, D. C. Carter, and F. Ruker. Conformational transitions of the three recombinant domains of human serum albumin depending on pH. J. Biol. Chem. 275:3042-3050 (2000).

    Google Scholar 

  29. C. Branden and J. Tooze. Introduction to Protein Structure, Garland Publishing, Inc., New York, 1991.

    Google Scholar 

  30. R. L. Stein. Catalysis by human leukocyte elastase: substrate structural dependence of rate-limiting protolytic catalysis and operation of the charge relay system. J. Am. Chem. Soc. 105:5111-5116 (1983).

    Google Scholar 

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Authors and Affiliations

  1. Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto, 862-0973, Japan

    Yuji Sakurai, Shen-Feng Ma, Hiroshi Watanabe & Masaki Otagiri

  2. The School of Pharmaceutical Sciences, Kitasato University, Tokyo, 108-8641, Japan

    Noriyuki Yamaotsu & Shuichi Hirono

  3. Faculty of Pharmaceutical Sciences, Nagoya City University, Nagoya, 467-8603, Japan

    Yukihisa Kurono

  4. Department of Medical Biochemistry, University of Aarhus, Aarhus C, DK-8000, Denmark

    Ulrich Kragh-Hansen

Authors
  1. Yuji Sakurai
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  2. Shen-Feng Ma
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  3. Hiroshi Watanabe
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  4. Noriyuki Yamaotsu
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  5. Shuichi Hirono
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  6. Yukihisa Kurono
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  7. Ulrich Kragh-Hansen
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  8. Masaki Otagiri
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Correspondence to Masaki Otagiri.

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Sakurai, Y., Ma, SF., Watanabe, H. et al. Esterase-Like Activity of Serum Albumin: Characterization of Its Structural Chemistry Using p-Nitrophenyl Esters as Substrates. Pharm Res 21, 285–292 (2004). https://doi.org/10.1023/B:PHAM.0000016241.84630.06

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