Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1991 Apr 1;275(Pt 1):159–164. doi: 10.1042/bj2750159

Heterogeneity of haem oxygenase 1 and 2 isoenzymes. Rat and primate transcripts for isoenzyme 2 differ in number and size.

G M Trakshel 1, J F Ewing 1, M D Maines 1
PMCID: PMC1150198  PMID: 2018471

Abstract

In Cebus apella monkey, as with other mammalian species tested to date, two different forms of haem oxygenase, HO-1 and HO-2, are detected. With the use of cDNA fragment corresponding to HO-1 nucleotides +71 to +833, blot hybridization of RNA revealed the presence of only one HO-1 mRNA of approx. 1.8 kb in both rat and monkey liver, kidney and brain. With the use of a full-length HO-2 DNA probe, blot hybridization of RNA isolated from the same rat organs revealed the presence of two HO-2 homologous transcripts of approx. 1.3 kb and approx. 1.9 kb. The same probe detected only one message of approx. 1.7 kb in monkey organs. The rat 1.3 kb mRNA has been previously shown [Rotenberg & Maines (1990) J. Biol. Chem. 265, 7501-7506] to encode HO-2 (36 kDa). The monkey 1.7 kb mRNA and the rat 1.3 kb mRNA encode proteins with similar molecular masses and immunochemical properties as indicated by Western-immunoblotting analysis. In rat organs the relative abundance of the two mRNAs differed as follows: in the liver the 1.3 kb mRNA was by far the most abundant form; in the brain equal amounts of the two mRNAs were detected, whereas in the kidney the 1.3 kb mRNA was somewhat more abundant. The protein encoded by the 1.8 kb HO-1 mRNA in the monkey did not exhibit immunochemical reactivity with antibody to rat HO-1 in Western blotting and direct e.l.i.s.a. analysis. The data suggest that, at the primary structural level, both HO-1 and HO-2 share extensive base sequence similarity in the rat and the Cebus apella monkey. The HO-1 protein, however, appears to undergo differential post-translational and/or conformational modifications in the two species, whereas the secondary structure of HO-2 protein and antigenic epitopes are conserved among the two mammalian species.

Full text

PDF
159

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Beale S. I., Cornejo J. Enzymatic heme oxygenase activity in soluble extracts of the unicellular red alga, Cyanidium caldarium. Arch Biochem Biophys. 1984 Dec;235(2):371–384. doi: 10.1016/0003-9861(84)90210-8. [DOI] [PubMed] [Google Scholar]
  2. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  3. Cruse I., Maines M. D. Evidence suggesting that the two forms of heme oxygenase are products of different genes. J Biol Chem. 1988 Mar 5;263(7):3348–3353. [PubMed] [Google Scholar]
  4. Dulley J. R., Grieve P. A. A simple technique for eliminating interference by detergents in the Lowry method of protein determination. Anal Biochem. 1975 Mar;64(1):136–141. doi: 10.1016/0003-2697(75)90415-7. [DOI] [PubMed] [Google Scholar]
  5. Engvall E. Enzyme immunoassay ELISA and EMIT. Methods Enzymol. 1980;70(A):419–439. doi: 10.1016/s0076-6879(80)70067-8. [DOI] [PubMed] [Google Scholar]
  6. Huang T. J., Trakshel G. M., Maines M. D. Detection of 10 variants of biliverdin reductase in rat liver by two-dimensional gel electrophoresis. J Biol Chem. 1989 May 15;264(14):7844–7849. [PubMed] [Google Scholar]
  7. Keyse S. M., Tyrrell R. M. Heme oxygenase is the major 32-kDa stress protein induced in human skin fibroblasts by UVA radiation, hydrogen peroxide, and sodium arsenite. Proc Natl Acad Sci U S A. 1989 Jan;86(1):99–103. doi: 10.1073/pnas.86.1.99. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kutty R. K., Maines M. D. Purification and characterization of biliverdin reductase from rat liver. J Biol Chem. 1981 Apr 25;256(8):3956–3962. [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Maines M. D. Heme oxygenase: function, multiplicity, regulatory mechanisms, and clinical applications. FASEB J. 1988 Jul;2(10):2557–2568. [PubMed] [Google Scholar]
  12. Maines M. D., Kappas A. Cobalt induction of hepatic heme oxygenase; with evidence that cytochrome P-450 is not essential for this enzyme activity. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4293–4297. doi: 10.1073/pnas.71.11.4293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Maines M. D., Trakshel G. M., Kutty R. K. Characterization of two constitutive forms of rat liver microsomal heme oxygenase. Only one molecular species of the enzyme is inducible. J Biol Chem. 1986 Jan 5;261(1):411–419. [PubMed] [Google Scholar]
  14. Rotenberg M. O., Maines M. D. Isolation, characterization, and expression in Escherichia coli of a cDNA encoding rat heme oxygenase-2. J Biol Chem. 1990 May 5;265(13):7501–7506. [PubMed] [Google Scholar]
  15. Shibahara S., Müller R. M., Taguchi H. Transcriptional control of rat heme oxygenase by heat shock. J Biol Chem. 1987 Sep 25;262(27):12889–12892. [PubMed] [Google Scholar]
  16. Shibahara S., Müller R., Taguchi H., Yoshida T. Cloning and expression of cDNA for rat heme oxygenase. Proc Natl Acad Sci U S A. 1985 Dec;82(23):7865–7869. doi: 10.1073/pnas.82.23.7865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Strobel H. W., Dignam J. D. Purification and properties of NADPH-cytochrome P-450 reductase. Methods Enzymol. 1978;52:89–96. doi: 10.1016/s0076-6879(78)52009-0. [DOI] [PubMed] [Google Scholar]
  18. Sun Y., Rotenberg M. O., Maines M. D. Developmental expression of heme oxygenase isozymes in rat brain. Two HO-2 mRNAs are detected. J Biol Chem. 1990 May 15;265(14):8212–8217. [PubMed] [Google Scholar]
  19. Tenhunen R., Marver H. S., Schmid R. The enzymatic conversion of heme to bilirubin by microsomal heme oxygenase. Proc Natl Acad Sci U S A. 1968 Oct;61(2):748–755. doi: 10.1073/pnas.61.2.748. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Trakshel G. M., Kutty R. K., Maines M. D. Purification and characterization of the major constitutive form of testicular heme oxygenase. The noninducible isoform. J Biol Chem. 1986 Aug 25;261(24):11131–11137. [PubMed] [Google Scholar]
  22. Trakshel G. M., Kutty R. K., Maines M. D. Resolution of the rat brain heme oxygenase activity: absence of a detectable amount of the inducible form (HO-1). Arch Biochem Biophys. 1988 Feb 1;260(2):732–739. doi: 10.1016/0003-9861(88)90503-6. [DOI] [PubMed] [Google Scholar]
  23. Trakshel G. M., Kutty R. K., Maines M. D. Resolution of the rat brain heme oxygenase activity: absence of a detectable amount of the inducible form (HO-1). Arch Biochem Biophys. 1988 Feb 1;260(2):732–739. doi: 10.1016/0003-9861(88)90503-6. [DOI] [PubMed] [Google Scholar]
  24. Trakshel G. M., Maines M. D. Detection of two heme oxygenase isoforms in the human testis. Biochem Biophys Res Commun. 1988 Jul 15;154(1):285–291. doi: 10.1016/0006-291x(88)90682-1. [DOI] [PubMed] [Google Scholar]
  25. Trakshel G. M., Maines M. D. Multiplicity of heme oxygenase isozymes. HO-1 and HO-2 are different molecular species in rat and rabbit. J Biol Chem. 1989 Jan 15;264(2):1323–1328. [PubMed] [Google Scholar]
  26. Yasukochi Y., Masters B. S. Some properties of a detergent-solubilized NADPH-cytochrome c(cytochrome P-450) reductase purified by biospecific affinity chromatography. J Biol Chem. 1976 Sep 10;251(17):5337–5344. [PubMed] [Google Scholar]
  27. Yoshida T., Biro P., Cohen T., Müller R. M., Shibahara S. Human heme oxygenase cDNA and induction of its mRNA by hemin. Eur J Biochem. 1988 Feb 1;171(3):457–461. doi: 10.1111/j.1432-1033.1988.tb13811.x. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES