Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1992 Sep 15;286(Pt 3):923–927. doi: 10.1042/bj2860923

Modification of vertebrate and algal prolyl 4-hydroxylases and vertebrate lysyl hydroxylase by diethyl pyrocarbonate. Evidence for histidine residues in the catalytic site of 2-oxoglutarate-coupled dioxygenases.

R Myllylä 1, V Günzler 1, K I Kivirikko 1, D D Kaska 1
PMCID: PMC1132991  PMID: 1329722

Abstract

A search for conserved amino acid residues within the cDNA-derived amino acid sequences of 2-oxoglutarate-coupled dioxygenases revealed the presence of two distinct motifs, spaced 49-71 amino acids apart, toward the C-terminal regions of these proteins. Each of the two common motifs contains an invariant histidine residue at a conserved position. The 2-oxoglutarate-coupled dioxygenases function in diverse processes, including the post-translational hydroxylation of proline and lysine residues in vertebrate collagens and the biosynthesis of microbial cephalosporins, yet they have a common reaction mechanisms, which requires the binding of Fe2+, 2-oxoglutarate, O2 and ascorbate at the catalytic site. The two regions of homology, and specifically the identical histidines, potentially represent functionally important sites related to their catalytic activity. Modification of histidine residues by diethyl pyrocarbonate inactivated vertebrate and algal prolyl 4-hydroxylase and vertebrate lysyl hydroxylase, indicating that histidine residues function in the catalytic site of these 2-oxoglutarate-coupled dioxygenases. Inactivation was prevented by the presence of co-substrates, but not by the peptide substrate. It is proposed that the histidine residues in the conserved motifs may function as Fe(2+)-binding ligands.

Full text

PDF
923

Selected References

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

  1. Bassuk J. A., Kao W. W., Herzer P., Kedersha N. L., Seyer J., DeMartino J. A., Daugherty B. L., Mark G. E., 3rd, Berg R. A. Prolyl 4-hydroxylase: molecular cloning and the primary structure of the alpha subunit from chicken embryo. Proc Natl Acad Sci U S A. 1989 Oct;86(19):7382–7386. doi: 10.1073/pnas.86.19.7382. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  3. Carr L. G., Skatrud P. L., Scheetz M. E., 2nd, Queener S. W., Ingolia T. D. Cloning and expression of the isopenicillin N synthetase gene from Penicillium chrysogenum. Gene. 1986;48(2-3):257–266. doi: 10.1016/0378-1119(86)90084-3. [DOI] [PubMed] [Google Scholar]
  4. De Jong L., Kemp A. Stoicheiometry and kinetics of the prolyl 4-hydroxylase partial reaction. Biochim Biophys Acta. 1984 May 31;787(1):105–111. doi: 10.1016/0167-4838(84)90113-4. [DOI] [PubMed] [Google Scholar]
  5. Deikman J., Fischer R. L. Interaction of a DNA binding factor with the 5'-flanking region of an ethylene-responsive fruit ripening gene from tomato. EMBO J. 1988 Nov;7(11):3315–3320. doi: 10.1002/j.1460-2075.1988.tb03202.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Günzler V., Hanauske-Abel H. M., Myllylä R., Kaska D. D., Hanauske A., Kivirikko K. I. Syncatalytic inactivation of prolyl 4-hydroxylase by anthracyclines. Biochem J. 1988 Apr 15;251(2):365–372. doi: 10.1042/bj2510365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hanauske-Abel H. M., Günzler V. A stereochemical concept for the catalytic mechanism of prolylhydroxylase: applicability to classification and design of inhibitors. J Theor Biol. 1982 Jan 21;94(2):421–455. doi: 10.1016/0022-5193(82)90320-4. [DOI] [PubMed] [Google Scholar]
  8. Hautala T., Byers M. G., Eddy R. L., Shows T. B., Kivirikko K. I., Myllylä R. Cloning of human lysyl hydroxylase: complete cDNA-derived amino acid sequence and assignment of the gene (PLOD) to chromosome 1p36.3----p36.2. Genomics. 1992 May;13(1):62–69. doi: 10.1016/0888-7543(92)90202-4. [DOI] [PubMed] [Google Scholar]
  9. Helaakoski T., Vuori K., Myllylä R., Kivirikko K. I., Pihlajaniemi T. Molecular cloning of the alpha-subunit of human prolyl 4-hydroxylase: the complete cDNA-derived amino acid sequence and evidence for alternative splicing of RNA transcripts. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4392–4396. doi: 10.1073/pnas.86.12.4392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Holdsworth M. J., Bird C. R., Ray J., Schuch W., Grierson D. Structure and expression of an ethylene-related mRNA from tomato. Nucleic Acids Res. 1987 Jan 26;15(2):731–739. doi: 10.1093/nar/15.2.731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kaska D. D., Günzler V., Kivirikko K. I., Myllylä R. Characterization of a low-relative-molecular-mass prolyl 4-hydroxylase from the green alga Chlamydomonas reinhardii. Biochem J. 1987 Jan 15;241(2):483–490. doi: 10.1042/bj2410483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kaska D. D., Myllylä R., Günzler V., Gibor A., Kivirikko K. I. Prolyl 4-hydroxylase from Volvox carteri. A low-Mr enzyme antigenically related to the alpha subunit of the vertebrate enzyme. Biochem J. 1988 Nov 15;256(1):257–263. doi: 10.1042/bj2560257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kivirikko K. I., Helaakoski T., Tasanen K., Vuori K., Myllylä R., Parkkonen T., Pihlajaniemi T. Molecular biology of prolyl 4-hydroxylase. Ann N Y Acad Sci. 1990;580:132–142. doi: 10.1111/j.1749-6632.1990.tb17925.x. [DOI] [PubMed] [Google Scholar]
  14. Kivirikko K. I., Myllylä R., Pihlajaniemi T. Protein hydroxylation: prolyl 4-hydroxylase, an enzyme with four cosubstrates and a multifunctional subunit. FASEB J. 1989 Mar;3(5):1609–1617. [PubMed] [Google Scholar]
  15. Kivirikko K. I., Myllylä R. Posttranslational enzymes in the biosynthesis of collagen: intracellular enzymes. Methods Enzymol. 1982;82(Pt A):245–304. doi: 10.1016/0076-6879(82)82067-3. [DOI] [PubMed] [Google Scholar]
  16. Kivirikko K. I., Myllylä R. Recent developments in posttranslational modification: intracellular processing. Methods Enzymol. 1987;144:96–114. doi: 10.1016/0076-6879(87)44175-x. [DOI] [PubMed] [Google Scholar]
  17. Koivu J., Myllylä R., Helaakoski T., Pihlajaniemi T., Tasanen K., Kivirikko K. I. A single polypeptide acts both as the beta subunit of prolyl 4-hydroxylase and as a protein disulfide-isomerase. J Biol Chem. 1987 May 15;262(14):6447–6449. [PubMed] [Google Scholar]
  18. Kovacevic S., Miller J. R. Cloning and sequencing of the beta-lactam hydroxylase gene (cefF) from Streptomyces clavuligerus: gene duplication may have led to separate hydroxylase and expandase activities in the actinomycetes. J Bacteriol. 1991 Jan;173(1):398–400. doi: 10.1128/jb.173.1.398-400.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kovacevic S., Weigel B. J., Tobin M. B., Ingolia T. D., Miller J. R. Cloning, characterization, and expression in Escherichia coli of the Streptomyces clavuligerus gene encoding deacetoxycephalosporin C synthetase. J Bacteriol. 1989 Feb;171(2):754–760. doi: 10.1128/jb.171.2.754-760.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Leskiw B. K., Aharonowitz Y., Mevarech M., Wolfe S., Vining L. C., Westlake D. W., Jensen S. E. Cloning and nucleotide sequence determination of the isopenicillin N synthetase gene from Streptomyces clavuligerus. Gene. 1988;62(2):187–196. doi: 10.1016/0378-1119(88)90557-4. [DOI] [PubMed] [Google Scholar]
  21. Majamaa K., Günzler V., Hanauske-Abel H. M., Myllylä R., Kivirikko K. I. Partial identity of the 2-oxoglutarate and ascorbate binding sites of prolyl 4-hydroxylase. J Biol Chem. 1986 Jun 15;261(17):7819–7823. [PubMed] [Google Scholar]
  22. Majamaa K., Hanauske-Abel H. M., Günzler V., Kivirikko K. I. The 2-oxoglutarate binding site of prolyl 4-hydroxylase. Identification of distinct subsites and evidence for 2-oxoglutarate decarboxylation in a ligand reaction at the enzyme-bound ferrous ion. Eur J Biochem. 1984 Jan 16;138(2):239–245. doi: 10.1111/j.1432-1033.1984.tb07907.x. [DOI] [PubMed] [Google Scholar]
  23. Matsuda J., Okabe S., Hashimoto T., Yamada Y. Molecular cloning of hyoscyamine 6 beta-hydroxylase, a 2-oxoglutarate-dependent dioxygenase, from cultured roots of Hyoscyamus niger. J Biol Chem. 1991 May 25;266(15):9460–9464. [PubMed] [Google Scholar]
  24. McGarvey D. J., Yu H., Christoffersen R. E. Nucleotide sequence of a ripening-related cDNA from avocado fruit. Plant Mol Biol. 1990 Jul;15(1):165–167. doi: 10.1007/BF00017736. [DOI] [PubMed] [Google Scholar]
  25. Miles E. W. Modification of histidyl residues in proteins by diethylpyrocarbonate. Methods Enzymol. 1977;47:431–442. doi: 10.1016/0076-6879(77)47043-5. [DOI] [PubMed] [Google Scholar]
  26. Myllylä R., Majamaa K., Günzler V., Hanauske-Abel H. M., Kivirikko K. I. Ascorbate is consumed stoichiometrically in the uncoupled reactions catalyzed by prolyl 4-hydroxylase and lysyl hydroxylase. J Biol Chem. 1984 May 10;259(9):5403–5405. [PubMed] [Google Scholar]
  27. Myllylä R., Pajunen L., Kivirikko K. I. Polyclonal and monoclonal antibodies to human lysyl hydroxylase and studies on the molecular heterogeneity of the enzyme. Biochem J. 1988 Jul 15;253(2):489–496. doi: 10.1042/bj2530489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Myllylä R., Pihlajaniemi T., Pajunen L., Turpeenniemi-Hujanen T., Kivirikko K. I. Molecular cloning of chick lysyl hydroxylase. Little homology in primary structure to the two types of subunit of prolyl 4-hydroxylase. J Biol Chem. 1991 Feb 15;266(5):2805–2810. [PubMed] [Google Scholar]
  29. Parkkonen T., Kivirikko K. I., Pihlajaniemi T. Molecular cloning of a multifunctional chicken protein acting as the prolyl 4-hydroxylase beta-subunit, protein disulphide-isomerase and a cellular thyroid-hormone-binding protein. Comparison of cDNA-deduced amino acid sequences with those in other species. Biochem J. 1988 Dec 15;256(3):1005–1011. doi: 10.1042/bj2561005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Pihlajaniemi T., Helaakoski T., Tasanen K., Myllylä R., Huhtala M. L., Koivu J., Kivirikko K. I. Molecular cloning of the beta-subunit of human prolyl 4-hydroxylase. This subunit and protein disulphide isomerase are products of the same gene. EMBO J. 1987 Mar;6(3):643–649. doi: 10.1002/j.1460-2075.1987.tb04803.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ramón D., Carramolino L., Patiño C., Sánchez F., Peñalva M. A. Cloning and characterization of the isopenicillin N synthetase gene mediating the formation of the beta-lactam ring in Aspergillus nidulans. Gene. 1987;57(2-3):171–181. doi: 10.1016/0378-1119(87)90120-x. [DOI] [PubMed] [Google Scholar]
  32. Salowe S. P., Marsh E. N., Townsend C. A. Purification and characterization of clavaminate synthase from Streptomyces clavuligerus: an unusual oxidative enzyme in natural product biosynthesis. Biochemistry. 1990 Jul 10;29(27):6499–6508. doi: 10.1021/bi00479a023. [DOI] [PubMed] [Google Scholar]
  33. Samson S. M., Belagaje R., Blankenship D. T., Chapman J. L., Perry D., Skatrud P. L., VanFrank R. M., Abraham E. P., Baldwin J. E., Queener S. W. Isolation, sequence determination and expression in Escherichia coli of the isopenicillin N synthetase gene from Cephalosporium acremonium. Nature. 1985 Nov 14;318(6042):191–194. doi: 10.1038/318191a0. [DOI] [PubMed] [Google Scholar]

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

RESOURCES