Skip to main content
SpringerLink
Log in

Modulation of the Activity of Newly Synthesized Human Phenylalanine Hydroxylase Mutant Proteins by Low-Molecular-Weight Compounds

  • Published:
The Protein Journal Aims and scope Submit manuscript

Abstract

Phenylketonuria, the most frequent disorder of amino acid metabolism, is caused by a deficient activity of human phenylalanine hydroxylase (hPAH). Rescue of the enzyme activity of several recombinant hPAH mutant forms (I65T, R261Q, R270K and V388M) by low molecular weight compounds namely glycerol, trimethylamine N-oxide (TMAO) and sodium 4-phenylbutyrate (4-PB) was investigated using a prokaryotic expression model. The studied compounds were added to the culture medium, in a concentration dependent manner, simultaneously to induction of protein expression. Among the tested molecules glycerol and TMAO were able to increase the enzyme activity of the studied mutant proteins. Furthermore, a decrease in aggregates and a recovery of the active tetrameric and dimeric forms were detected. Since the addition of the studied compounds to the medium did not change the expression level of E. Coli molecular chaperones we postulate that glycerol and TMAO rescue results from a direct stabilizing effect of the newly synthesized mutant hPAH enzymes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

BH4 :

(R)-L-erythro-5,6,7,8-tetrahydrobiopterin

PKU:

Phenylketonuria

HPA:

Hyperphenylalaninemia

MWCO:

Molecular weight cut-off

SEC:

Size exclusion chromatography

HRP:

Horseradish peroxidase

References

  1. Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1994) Current protocols in molecular biology. John Wiley and Sons, New York

    Google Scholar 

  2. Baskakov I, Wang A, Bolen DW (1998) Biophys J 74:2666–2673

    Article  CAS  Google Scholar 

  3. Bjorgo E, de Carvalho RM, Flatmark T (2001) Eur J Biochem 268:997–1005

    Article  CAS  Google Scholar 

  4. Bolen DW, Baskakov IV (2001) J Mol Biol 310:955–963

    Article  CAS  Google Scholar 

  5. Bradford MM (1976) Anal Biochem 72:248–254

    Article  CAS  Google Scholar 

  6. Burrows JA, Willis LK, Perlmutter DH (2000) Proc Natl Acad Sci U S A 97:1796–1801

    Article  CAS  Google Scholar 

  7. Butler SL, Falke JJ (1996) Biochemistry 35:10595–10600

    Article  CAS  Google Scholar 

  8. da Costa MS, Santos H, Galinski EA (1998) Adv Biochem Eng Biotechnol 61:117–153

    Google Scholar 

  9. Davis MD, Parniak MA, Kaufman S, Kempner E (1996) Arch Biochem Biophys 325:235–241

    Article  CAS  Google Scholar 

  10. de Almeida SF, Picarote G, Fleming JV, Carmo-Fonseca M, Azevedo JE, de Sousa M (2007) J Biol Chem 282:27905–27912

    Article  CAS  Google Scholar 

  11. Gamez A, Perez B, Ugarte M, Desviat LR (2000) J Biol Chem 275:29737–29742

    Article  CAS  Google Scholar 

  12. Hansen PA, Waheed A, Corbett JA (2007) Trends Endocrinol Metab 18:1–3

    Article  CAS  Google Scholar 

  13. Kaufman S (1987) Methods Enzymol 142:3–17

    Article  CAS  Google Scholar 

  14. Knappskog PM, Flatmark T, Aarden JM, Haavik J, Martinez A (1996) Eur J Biochem 242:813–821

    Article  CAS  Google Scholar 

  15. Kowlessur D, Citron BA, Kaufman S (1996) Arch Biochem Biophys 333:85–95

    Article  CAS  Google Scholar 

  16. Kumar R, Serrette JM, Thompson EB (2005) Arch Biochem Biophys 436:78–82

    Article  CAS  Google Scholar 

  17. Kwok SC, Ledley FD, DiLella AG, Robson KJ, Woo SL (1985) Biochemistry 24:556–561

    Article  CAS  Google Scholar 

  18. Leandro P, Rivera I, Lechner MC, de Almeida IT, Konecki D (2000) Mol Genet Metab 69:204–212

    Article  CAS  Google Scholar 

  19. Leandro P, Lechner MC, Tavares de Almeida I, Konecki D (2001) Mol Genet Metab 73:173–178

    Article  CAS  Google Scholar 

  20. Leandro P, Rivera I, Lechner MC, Konecki D, Almeida I (2001) Gene Function and Disease 2:46–50

    Article  CAS  Google Scholar 

  21. Martinez A, Knappskog PM, Olafsdottir S, Doskeland AP, Eiken HG, Svebak RM, Bozzini M, Apold J, Flatmark T (1995) Biochem J 306(Pt 2):589–597

    CAS  Google Scholar 

  22. Perlmutter DH (2002) Pediatr Res 52:832–836

    Google Scholar 

  23. Pey AL, Stricher F, Serrano L, Martinez A (2007) Am J Hum Genet 81:1006–1024

    Article  CAS  Google Scholar 

  24. Pradeep L, Udgaonkar JB (2004) J Biol Chem 279:40303–40313

    Article  CAS  Google Scholar 

  25. Rivera I, Cabral A, Almeida M, Leandro P, Carmona C, Eusebio F, Tasso T, Vilarinho L, Martins E, Lechner MC, de Almeida IT, Konecki DS, Lichter-Konecki U (2000) Mol Genet Metab 69:195–203

    Article  CAS  Google Scholar 

  26. Saudubray JM, Sedel F, Walter JH (2006) J Inherit Metab Dis 29:261–274

    Article  Google Scholar 

  27. Scriver CR (2007) Hum Mutat 28:831–845

    Article  CAS  Google Scholar 

  28. Simon LM, Kotorman M, Garab G, Laczko I (2002) Biochem Biophys Res Commun 293:416–420

    Article  CAS  Google Scholar 

  29. Singh LR, Chen X, Kozich V, Kruger WD (2007) Mol Genet Metab 91:335–342

    Article  CAS  Google Scholar 

  30. Song JL, Chuang DT (2001) J Biol Chem 276:40241–40246

    CAS  Google Scholar 

  31. Stokka AJ, Flatmark T (2003) Biochem J 369:509–518

    Article  CAS  Google Scholar 

  32. Uversky VN, Li J, Fink AL (2001) FEBS Lett 509:31–35

    Article  CAS  Google Scholar 

Download references

Acknowledgements

F. Ventura (iMed.UL) is gratefully acknowledged for critically reading of the manuscript. This work was supported by Fundação para a Ciência e Tecnologia, Portugal, and FEDER (MGI/POCTI/40844/2001, SFRH/BD/10807/2002 and SFRH/BD/19024/2004).

Author information

Authors and Affiliations

  1. Metabolism and Genetics Group, iMed.UL, Faculdade Farmácia da Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003, Lisbon, Portugal

    Cátia Nascimento, João Leandro, Isabel Tavares de Almeida & Paula Leandro

Authors
  1. Cátia Nascimento
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. João Leandro
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Isabel Tavares de Almeida
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Paula Leandro
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Paula Leandro.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nascimento, C., Leandro, J., Tavares de Almeida, I. et al. Modulation of the Activity of Newly Synthesized Human Phenylalanine Hydroxylase Mutant Proteins by Low-Molecular-Weight Compounds. Protein J 27, 392–400 (2008). https://doi.org/10.1007/s10930-008-9149-9

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10930-008-9149-9

Keywords

Search

Navigation