Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1992 Aug;174(16):5391–5399. doi: 10.1128/jb.174.16.5391-5399.1992

Architecture of peroxisomal alcohol oxidase crystals from the methylotrophic yeast Hansenula polymorpha as deduced by electron microscopy.

J Vonck 1, E F van Bruggen 1
PMCID: PMC206377  PMID: 1644766

Abstract

The architecture of alcohol oxidase crystalloids occurring in vivo in the peroxisomes of methylotrophic yeasts was deduced from electron micrographs of similar crystals of the Hansenula polymorpha enzyme grown in vitro. Three characteristic views of the crystal are observed, as well as single layers in the very early stages of crystal formation. The crystal is concluded to be cubical, with every octameric molecule making the same contacts with four neighbors in one plane, at right angles to its fourfold axis. The unit cell contains six octamers, in three mutually orthogonal orientations, and two large holes, which can accommodate other peroxisomal proteins involved in methanol metabolism. The crystal contains channels, connecting the holes, which allow the diffusion of relatively large molecules through the crystal. Crystal formation depends on just one contact per subunit, which may explain the fragility of the crystals.

Full text

PDF
5391

Images in this article

Selected References

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

  1. Boys C. W., Hill D. J., Stockley P. G., Woodward J. R. Crystallization of alcohol oxidase from Pichia pastoris. J Mol Biol. 1989 Jul 5;208(1):211–212. doi: 10.1016/0022-2836(89)90099-5. [DOI] [PubMed] [Google Scholar]
  2. Goodman J. M., Scott C. W., Donahue P. N., Atherton J. P. Alcohol oxidase assembles post-translationally into the peroxisome of Candida boidinii. J Biol Chem. 1984 Jul 10;259(13):8485–8493. [PubMed] [Google Scholar]
  3. Janowicz Z. A., Eckart M. R., Drewke C., Roggenkamp R. O., Hollenberg C. P., Maat J., Ledeboer A. M., Visser C., Verrips C. T. Cloning and characterization of the DAS gene encoding the major methanol assimilatory enzyme from the methylotrophic yeast Hansenula polymorpha. Nucleic Acids Res. 1985 May 10;13(9):3043–3062. doi: 10.1093/nar/13.9.3043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Kato N., Omori Y., Tani Y., Ogata K. Alcohol oxidases of Kloeckera sp. and Hansenula polymorpha. Catalytic properties and subunit structures. Eur J Biochem. 1976 May 1;64(2):341–350. doi: 10.1111/j.1432-1033.1976.tb10307.x. [DOI] [PubMed] [Google Scholar]
  5. Ledeboer A. M., Edens L., Maat J., Visser C., Bos J. W., Verrips C. T., Janowicz Z., Eckart M., Roggenkamp R., Hollenberg C. P. Molecular cloning and characterization of a gene coding for methanol oxidase in Hansenula polymorpha. Nucleic Acids Res. 1985 May 10;13(9):3063–3082. doi: 10.1093/nar/13.9.3063. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Murthy M. R., Reid T. J., 3rd, Sicignano A., Tanaka N., Rossmann M. G. Structure of beef liver catalase. J Mol Biol. 1981 Oct 25;152(2):465–499. doi: 10.1016/0022-2836(81)90254-0. [DOI] [PubMed] [Google Scholar]
  7. Osumi M., Nagano M., Yamada N., Hosoi J., Yanagida M. Three-dimensional structure of the crystalloid in the microbody of Kloeckera sp.: composite crystal model. J Bacteriol. 1982 Jul;151(1):376–383. doi: 10.1128/jb.151.1.376-383.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Roa M., Blobel G. Biosynthesis of peroxisomal enzymes in the methylotrophic yeast Hansenula polymorpha. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6872–6876. doi: 10.1073/pnas.80.22.6872. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Sahm H., Wagner F. Microbial assimilation of methanol. The ethanol- and methanol-oxidizing enzymes of the yeast Candida boidinii. Eur J Biochem. 1973 Jul 2;36(1):250–256. doi: 10.1111/j.1432-1033.1973.tb02907.x. [DOI] [PubMed] [Google Scholar]
  10. Seah T. C., Kaplan J. G. Purification and properties of the catalase of bakers' yeast. J Biol Chem. 1973 Apr 25;248(8):2889–2893. [PubMed] [Google Scholar]
  11. Tykarska E., Lebioda L., Marchut E., Steczko J., Stec B. Crystallization of alcohol oxidase from Pichia pastoris. Secondary structure predictions indicate a domain with the eightfold beta/alpha-barrel fold. J Protein Chem. 1990 Feb;9(1):83–86. doi: 10.1007/BF01024988. [DOI] [PubMed] [Google Scholar]
  12. Van der Klei I. J., Lawson C. L., Rozeboom H., Dijkstra B. W., Veenhuis M., Harder W., Hol W. G. Use of electron microscopy in the examination of lattice defects in crystals of alcohol oxidase. FEBS Lett. 1989 Feb 13;244(1):213–216. doi: 10.1016/0014-5793(89)81195-0. [DOI] [PubMed] [Google Scholar]
  13. Veenhuis M., Harder W., van Dijken J. P., Mayer F. Substructure of crystalline peroxisomes in methanol-grown Hansenula polymorpha: evidence for an in vivo crystal of alcohol oxidase. Mol Cell Biol. 1981 Oct;1(10):949–957. doi: 10.1128/mcb.1.10.949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Veenhuis M., van Dijken J. P., Pilon S. A., Harder W. Development of crystalline peroxisomes in methanol-grown cells of the yeast Hansenula polymorpha and its relation to environmental conditions. Arch Microbiol. 1978 May 30;117(2):153–163. doi: 10.1007/BF00402303. [DOI] [PubMed] [Google Scholar]
  15. Vonck J., van Bruggen E. F. Electron microscopy and image analysis of two-dimensional crystals and single molecules of alcohol oxidase from Hansenula polymorpha. Biochim Biophys Acta. 1990 Mar 29;1038(1):74–79. doi: 10.1016/0167-4838(90)90012-5. [DOI] [PubMed] [Google Scholar]
  16. van der Klei I. J., Bystrykh L. V., Harder W. Alcohol oxidase from Hansenula polymorpha CBS 4732. Methods Enzymol. 1990;188:420–427. doi: 10.1016/0076-6879(90)88067-k. [DOI] [PubMed] [Google Scholar]
  17. van der Klei I. J., Harder W., Veenhuis M. Biosynthesis and assembly of alcohol oxidase, a peroxisomal matrix protein in methylotrophic yeasts: a review. Yeast. 1991 Apr;7(3):195–209. doi: 10.1002/yea.320070302. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES