Skip to main content
Protein Science : A Publication of the Protein Society logoLink to Protein Science : A Publication of the Protein Society
. 2000 Mar;9(3):544–551. doi: 10.1110/ps.9.3.544

The X-ray structure of a chitinase from the pathogenic fungus Coccidioides immitis.

T Hollis 1, A F Monzingo 1, K Bortone 1, S Ernst 1, R Cox 1, J D Robertus 1
PMCID: PMC2144563  PMID: 10752616

Abstract

The X-ray structure of chitinase from the fungal pathogen Coccidioides immitis has been solved to 2.2 A resolution. Like other members of the class 18 hydrolase family, this 427 residue protein is an eight-stranded beta/alpha-barrel. Although lacking an N-terminal chitin anchoring domain, the enzyme closely resembles the chitinase from Serratia marcescens. Among the conserved features are three cis peptide bonds, all involving conserved active site residues. The active site is formed from conserved residues such as tryptophans 47, 131, 315, 378, tyrosines 239 and 293, and arginines 52 and 295. Glu171 is the catalytic acid in the hydrolytic mechanism; it was mutated to a Gln, and activity was abolished. Allosamidin is a substrate analog that strongly inhibits the class 18 enzymes. Its binding to the chitinase hevamine has been observed, and we used conserved structural features of the two enzymes to predict the inhibitors binding to the fungal enzyme.

Full Text

The Full Text of this article is available as a PDF (690.2 KB).

Selected References

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

  1. Ampel N. M. Emerging disease issues and fungal pathogens associated with HIV infection. Emerg Infect Dis. 1996 Apr-Jun;2(2):109–116. doi: 10.3201/eid0202.960205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Armand S., Tomita H., Heyraud A., Gey C., Watanabe T., Henrissat B. Stereochemical course of the hydrolysis reaction catalyzed by chitinases A1 and D from Bacillus circulans WL-12. FEBS Lett. 1994 Apr 25;343(2):177–180. doi: 10.1016/0014-5793(94)80314-5. [DOI] [PubMed] [Google Scholar]
  3. Blaiseau P. L., Lafay J. F. Primary structure of a chitinase-encoding gene (chi1) from the filamentous fungus Aphanocladium album: similarity to bacterial chitinases. Gene. 1992 Oct 21;120(2):243–248. doi: 10.1016/0378-1119(92)90099-b. [DOI] [PubMed] [Google Scholar]
  4. Blake C. C., Johnson L. N., Mair G. A., North A. C., Phillips D. C., Sarma V. R. Crystallographic studies of the activity of hen egg-white lysozyme. Proc R Soc Lond B Biol Sci. 1967 Apr 18;167(1009):378–388. doi: 10.1098/rspb.1967.0035. [DOI] [PubMed] [Google Scholar]
  5. Brameld K. A., Shrader W. D., Imperiali B., Goddard W. A., 3rd Substrate assistance in the mechanism of family 18 chitinases: theoretical studies of potential intermediates and inhibitors. J Mol Biol. 1998 Jul 31;280(5):913–923. doi: 10.1006/jmbi.1998.1890. [DOI] [PubMed] [Google Scholar]
  6. Bulawa C. E., Osmond B. C. Chitin synthase I and chitin synthase II are not required for chitin synthesis in vivo in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7424–7428. doi: 10.1073/pnas.87.19.7424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dolan M. J., Cox R. A. Production and characterization of a monoclonal antibody to the complement fixation antigen of Coccidioides immitis. Infect Immun. 1991 Jun;59(6):2175–2180. doi: 10.1128/iai.59.6.2175-2180.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. García I., Lora J. M., de la Cruz J., Benítez T., Llobell A., Pintor-Toro J. A. Cloning and characterization of a chitinase (chit42) cDNA from the mycoparasitic fungus Trichoderma harzianum. Curr Genet. 1994 Dec;27(1):83–89. doi: 10.1007/BF00326583. [DOI] [PubMed] [Google Scholar]
  9. Hart P. J., Pfluger H. D., Monzingo A. F., Hollis T., Robertus J. D. The refined crystal structure of an endochitinase from Hordeum vulgare L. seeds at 1.8 A resolution. J Mol Biol. 1995 Apr 28;248(2):402–413. [PubMed] [Google Scholar]
  10. Henrissat B., Bairoch A. New families in the classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem J. 1993 Aug 1;293(Pt 3):781–788. doi: 10.1042/bj2930781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Higgins D. G., Bleasby A. J., Fuchs R. CLUSTAL V: improved software for multiple sequence alignment. Comput Appl Biosci. 1992 Apr;8(2):189–191. doi: 10.1093/bioinformatics/8.2.189. [DOI] [PubMed] [Google Scholar]
  12. Hollis T., Monzingo A. F., Bortone K., Schelp E., Cox R., Robertus J. D. Crystallization and preliminary X-ray analysis of a chitinase from the fungal pathogen Coccidioides immitis. Acta Crystallogr D Biol Crystallogr. 1998 Nov 1;54(Pt 6 2):1412–1413. doi: 10.1107/s0907444998008531. [DOI] [PubMed] [Google Scholar]
  13. Johnson S. M., Pappagianis D. The coccidioidal complement fixation and immunodiffusion-complement fixation antigen is a chitinase. Infect Immun. 1992 Jul;60(7):2588–2592. doi: 10.1128/iai.60.7.2588-2592.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jones J. D., Grady K. L., Suslow T. V., Bedbrook J. R. Isolation and characterization of genes encoding two chitinase enzymes from Serratia marcescens. EMBO J. 1986 Mar;5(3):467–473. doi: 10.1002/j.1460-2075.1986.tb04235.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jones T. A., Zou J. Y., Cowan S. W., Kjeldgaard M. Improved methods for building protein models in electron density maps and the location of errors in these models. Acta Crystallogr A. 1991 Mar 1;47(Pt 2):110–119. doi: 10.1107/s0108767390010224. [DOI] [PubMed] [Google Scholar]
  16. Kleywegt G. J., Jones T. A. xdlMAPMAN and xdlDATAMAN - programs for reformatting, analysis and manipulation of biomacromolecular electron-density maps and reflection data sets. Acta Crystallogr D Biol Crystallogr. 1996 Jul 1;52(Pt 4):826–828. doi: 10.1107/S0907444995014983. [DOI] [PubMed] [Google Scholar]
  17. Kuranda M. J., Robbins P. W. Chitinase is required for cell separation during growth of Saccharomyces cerevisiae. J Biol Chem. 1991 Oct 15;266(29):19758–19767. [PubMed] [Google Scholar]
  18. Milewski S., O'Donnell R. W., Gooday G. W. Chemical modification studies of the active centre of Candida albicans chitinase and its inhibition by allosamidin. J Gen Microbiol. 1992 Dec;138(12):2545–2550. doi: 10.1099/00221287-138-12-2545. [DOI] [PubMed] [Google Scholar]
  19. Minamoto G. Y., Rosenberg A. S. Fungal infections in patients with acquired immunodeficiency syndrome. Med Clin North Am. 1997 Mar;81(2):381–409. doi: 10.1016/s0025-7125(05)70523-x. [DOI] [PubMed] [Google Scholar]
  20. Monzingo A. F., Marcotte E. M., Hart P. J., Robertus J. D. Chitinases, chitosanases, and lysozymes can be divided into procaryotic and eucaryotic families sharing a conserved core. Nat Struct Biol. 1996 Feb;3(2):133–140. doi: 10.1038/nsb0296-133. [DOI] [PubMed] [Google Scholar]
  21. Nishimoto Y., Sakuda S., Takayama S., Yamada Y. Isolation and characterization of new allosamidins. J Antibiot (Tokyo) 1991 Jul;44(7):716–722. doi: 10.7164/antibiotics.44.716. [DOI] [PubMed] [Google Scholar]
  22. Pappagianis D., Zimmer B. L. Serology of coccidioidomycosis. Clin Microbiol Rev. 1990 Jul;3(3):247–268. doi: 10.1128/cmr.3.3.247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Perrakis A., Tews I., Dauter Z., Oppenheim A. B., Chet I., Wilson K. S., Vorgias C. E. Crystal structure of a bacterial chitinase at 2.3 A resolution. Structure. 1994 Dec 15;2(12):1169–1180. doi: 10.1016/s0969-2126(94)00119-7. [DOI] [PubMed] [Google Scholar]
  24. Pishko E. J., Kirkland T. N., Cole G. T. Isolation and characterization of two chitinase-encoding genes (cts1, cts2) from the fungus Coccidioides immitis. Gene. 1995 Dec 29;167(1-2):173–177. doi: 10.1016/0378-1119(95)00654-0. [DOI] [PubMed] [Google Scholar]
  25. Terwisscha van Scheltinga A. C., Armand S., Kalk K. H., Isogai A., Henrissat B., Dijkstra B. W. Stereochemistry of chitin hydrolysis by a plant chitinase/lysozyme and X-ray structure of a complex with allosamidin: evidence for substrate assisted catalysis. Biochemistry. 1995 Dec 5;34(48):15619–15623. doi: 10.1021/bi00048a003. [DOI] [PubMed] [Google Scholar]
  26. Terwisscha van Scheltinga A. C., Hennig M., Dijkstra B. W. The 1.8 A resolution structure of hevamine, a plant chitinase/lysozyme, and analysis of the conserved sequence and structure motifs of glycosyl hydrolase family 18. J Mol Biol. 1996 Sep 20;262(2):243–257. doi: 10.1006/jmbi.1996.0510. [DOI] [PubMed] [Google Scholar]
  27. Watanabe T., Kobori K., Miyashita K., Fujii T., Sakai H., Uchida M., Tanaka H. Identification of glutamic acid 204 and aspartic acid 200 in chitinase A1 of Bacillus circulans WL-12 as essential residues for chitinase activity. J Biol Chem. 1993 Sep 5;268(25):18567–18572. [PubMed] [Google Scholar]
  28. Yang C., Zhu Y., Magee D. M., Cox R. A. Molecular cloning and characterization of the Coccidioides immitis complement fixation/chitinase antigen. Infect Immun. 1996 Jun;64(6):1992–1997. doi: 10.1128/iai.64.6.1992-1997.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Protein Science : A Publication of the Protein Society are provided here courtesy of The Protein Society

RESOURCES