Skip to main content
SpringerLink
Log in

Biochemical Characterization of Porphobilinogen Deaminase-Deficient Mice During Phenobarbital Induction of Heme Synthesis and the Effect of Enzyme Replacement

  • Articles
  • Published:
Molecular Medicine Aims and scope Submit manuscript

Abstract

Acute intermittent porphyria (AIP) is a genetic disorder caused by a deficiency of porphobilinogen deaminase (PBGD), the 3rd enzyme in heme synthesis. It is clinically characterized by acute attacks of neuropsychiatric symptoms and biochemically by increased urinary excretion of the porphyrin precursors porphobilinogen (PBG) and 5-aminolevulinic acid (ALA). A mouse model that is partially deficient in PBGD and biochemically mimics AIP after induction of the hepatic ALA synthase by phenobarbital was used in this study to identify the site of formation of the presumably toxic porphyrin precursors and study the effect of enzyme-replacement therapy by using recombinant human PBGD (rhPBGD). After 4 d of phenobarbital administration, high levels of PBG and ALA were found in liver, kidney, plasma, and urine of the PBGD-deficient mice. The administration of rhPBGD intravenously or subcutaneously after a 4-d phenobarbital induction was shown to lower the PBG level in plasma in a dose-dependent manner with maximal effect seen after 30 min and 2 h, respectively. Injection of rhPBGD subcutaneously twice daily during a 4-d phenobarbital induction reduced urinary PBG excretion to 25% of the levels found in PBGD-deficient mice administered with only phenobarbital. This study points to the liver as the main producer of PBG and ALA in the phenobarbital-induced PBGD-deficient mice and demonstrates efficient removal of accumulated PBG in plasma and urine by enzyme-replacement therapy.

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

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

References

  1. Anderson KE, Sassa S, Bishop DF, Desnick RJ. (2001) Disorders of Heme Biosynthesis: X-Linked Sideroblastic Anemia and the Porphyrias. In: The Metabolic and Molecular Bases of Inherited Disease (Vol. 2). Scriver CR, Beaudet AL, Valle D, Sly WS (eds.). McGraw-Hill, New York, USA, pp. 2991–3062.

    Google Scholar 

  2. Granick S. (1966) The induction in vitro of the synthesis of delta-aminolevulinic acid synthetase in chemical porphyria: A response to certain drugs, sex hormones, and foreign chemicals. J. Biol. Chem. 241:1359–75.

    CAS  PubMed  Google Scholar 

  3. Miyagi K, Cardinal R, Bossenmaier I, Watson CJ. (1971) The serum porphobilinogen and hepatic porphobilinogen deaminase in normal and porphyric individuals. J. Lab. Clin. Med. 78(5):683–95.

    CAS  PubMed  Google Scholar 

  4. Schmid R, Schwartz S, Watson CJ. (1954) Porphyrin content of bone marrow and liver in the various forms of porphyria. AMA Arch. Int. Med. 93:167–90.

    Article  CAS  Google Scholar 

  5. Sweeney VP, Pathak MA, Asbury AK. (1970) Acute intermittent porphyria. Increased ALA-synthetase activity during an acute attack. Brain. 93:369–80.

    Article  CAS  Google Scholar 

  6. Goldberg A, Rimington C. (1954) Fate of porphobilinogen in the rat. Relation to acute porphyria in man. Lancet 2:172–3.

    Article  Google Scholar 

  7. Thunell S. (2000) Porphyrins, porphyrin metabolism and porphyrias. I. Update. Scand. J. Clin. Lab. Invest. 60(7):509–40.

    Article  CAS  Google Scholar 

  8. Bechara EJH. (1996) Oxidative stress in acute intermittent porphyria and lead poisoning may be triggered by 5-aminolevulinic acid. Braz. J. Med. Biol. Res. 29:841–51.

    CAS  PubMed  Google Scholar 

  9. Kauppinen R, Timonen K, Mustajoki P. (1994) Treatment of the porphyrias. Ann. Med. 26:31–8.

    Article  CAS  Google Scholar 

  10. Tenhunen R, Mustajoki P. (1998) Acute porphyria: treatment with heme. Semin. Liver Dis. 18(1):53–5.

    Article  CAS  Google Scholar 

  11. Badminton MN, Elder GH. (2002) Management of acute and cutaneous porphyrias. Int. J. Clin. Pract. 56(4):272–8.

    CAS  PubMed  Google Scholar 

  12. Sardh E, Rejkjaer L, Harper P, Andersson DE. (2003) First clinical trial of i.v. rhPBGD in healthy subjects with and without diagnosed manifest acute intermittent porphyria (AIP). Physiol. Res. 52:23S.

    Google Scholar 

  13. Grandchamp B, De Verneuil H, Beaumont C, Chretien S, Walter O, Nordmann Y. (1987) Tissue-specific expression of porphobilinogen deaminase. Two isoenzymes from a single gene. Eur. J. Biochem. 162(1):105–10.

    Article  CAS  Google Scholar 

  14. Lindberg RL et al. (1996) Porphobilinogen deaminase deficiency in mice causes a neuropathy resembling that of human hepatic porphyria. Nat Genet. 12(2):195–9.

    Article  CAS  Google Scholar 

  15. Jover R, Hoffmann F, Scheffler-Koch V, Lindberg RL. (2000) Limited heme synthesis in porphobilinogen deaminase-deficient mice impairs transcriptional activation of specific cytochrome P450 genes by phenobarbital. Eur. J. Biochem. 267(24):7128–37.

    Article  CAS  Google Scholar 

  16. Magnussen CR, Levine JB, Doherty JM, Cheesman JO, Tschudy DP (1974). A red cell enzyme method for the diagnosis of acute intermittent porphyria. Blood. 44(6):857–68.

    CAS  PubMed  Google Scholar 

  17. Johansson A, Möller C, Gellerfors P, Harper P. (2002) Non-viral mediated gene transfer of porphobilinogen deaminase into mammalian cells. Scand. J. Clin. Lab. Invest. 62(2):105–14.

    Article  CAS  Google Scholar 

  18. Lentner C. (1984) Physical chemistry composition of blood hematology somatometric data. In: Geigy Scientific Tables (Vol. 3),Ciba-Geigy, pp. 166–7.

    Google Scholar 

  19. Davis JR, Andelman SL. (1967) Urinary delta-aminolevulinic acid (ALA) levels in lead poisoning. I. A modified method for the rapid determination of urinary delta-aminolevulinic acid using disposable ion-exchange chromatography columns. Arch. Envir. Health. 15:53–9.

    Article  CAS  Google Scholar 

  20. Bannerman RM. (1983) Hematology. In: The mouse in medical research (Vol. III). Foster HL, Small JD, Fox JG (eds.). Academic Press, Orlando, FL, USA, pp. 293–312.

    Article  Google Scholar 

  21. Sassa S. (1976) Sequential induction of heme pathway enzymes during erythroid differentiation of mouse friend leukemia virus-infected cells. J. Exp. Med. 143:305–15.

    Article  CAS  Google Scholar 

  22. Lindberg RL et al. (1999) Motor neuropathy in porphobilinogen deaminase-deficient mice imitates the peripheral neuropathy of human acute porphyria. J. Clin. Invest. 103(8):1127–34.

    Article  CAS  Google Scholar 

  23. Harper P, Floderus Y, Andersson C, Möller C, Rejkjaer L, Sardh E, Andersson DE. (2003) Correlation between plasma and urinary levels of porphobilinogen (PBG) and 5-aminolevulinic acid (ALA) in ten asymptomatic gene carriers of acute intermittent porphyria (AIP) with increased porphyrin precursor excretion. Physiol. Res. 52:10S.

    Google Scholar 

  24. Bossenmaier I, Cardinal R. (1968) Stability of delta-aminolevulinic acid and porphobilinogen in urine under varying conditions. Clin. Chem. 14:610–4.

    CAS  PubMed  Google Scholar 

  25. Elder GH. (1998) Genetic defects in the porphyrias: types and significance. Clin. Dermatol. 16(2):225–33.

    Article  CAS  Google Scholar 

  26. Thunell S et al. (1995) Markers for vulnerability in acute porphyria. A hypothesis paper. Eur. J. Clin. Chem. Clin. Biochem. 33(4):179–94.

    CAS  PubMed  Google Scholar 

  27. Dowdle E, Mustard P, Spong N, Eales L. (1968) The metabolism of [5-14C]delta-aminolaevulinic acid in normal and porphyric human subjects. Clin. Sci. 34: 233–51.

    CAS  PubMed  Google Scholar 

  28. Mustajoki P, Timonen K, Gorchein A, Seppalainen AM, Matikainen E, Tenhunen R. (1992) Sustained high plasma 5-aminolaevulinic acid concentration in a volunteer: no porphyric symptoms. Eur. J. Clin. Invest. 22(6):407–11.

    Article  CAS  Google Scholar 

  29. Edwards SR, Shanley BC, Reynoldson JA. (1984) Neuropharmacology of delta-aminolaevulinic acid-I. Effect of acute administration in rodents. Neuropharmacology 23:477–81.

    Article  CAS  Google Scholar 

  30. Feldman DS, Levere RD, Lieberman JS, Cardinal RA, Watson CJ. (1971) Presynaptic neuromuscular inhibition by porphobilinogen and porphobilin. Proc. Nat. Acad. Sci. U.S.A. 68:383–6.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We wish to thank Associate Professor Stig Thunell for reviewing the manuscript, and Anette Pettersson and Helena Reuterwall at HemeBiotech A/S for technical assistance with the LC-MS method and Western blot. The work was supported by grants from Clas Groschinsky’s Memorial Fund and the Karolinska Institute.

Author information

Authors and Affiliations

  1. Department of Laboratory Medicine, Division of Clinical Chemistry, Karolinska Institute, Karolinska University Hospital, Huddinge, Porphyria Centre Sweden, Centre for Metabolic Inherited Diseases (CMMS) C2 71, Stockholm, 141 86, Sweden

    Annika Johansson & Pauline Harper

  2. HemeBiotech A/S, Lidingö, Sweden

    Christer Möller & Jens Fogh

Authors
  1. Annika Johansson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Christer Möller
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jens Fogh
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Pauline Harper
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Annika Johansson.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Johansson, A., Möller, C., Fogh, J. et al. Biochemical Characterization of Porphobilinogen Deaminase-Deficient Mice During Phenobarbital Induction of Heme Synthesis and the Effect of Enzyme Replacement. Mol Med 9, 193–199 (2003). https://doi.org/10.2119/2004-00002.Johansson

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.2119/2004-00002.Johansson

Keywords

Search

Navigation