Crystallization and Preliminary X-ray Analysis of a Family 19 Glycosyl Hydrolase from Carica Papaya Latex

Overview

Journal Acta Crystallogr Sect F Struct Biol Cryst Commun

Date 2008 May 6

PMID 18453704

Citations 3

Authors

Joelle Huet

Mohamed Azarkan

Yvan Looze

Vincent Villeret

Rene Wintjens

Affiliations

Soon will be listed here.

Abstract

A chitinase isolated from the latex of the tropical species Carica papaya has been purified to homogeneity and crystallized. This enzyme belongs to glycosyl hydrolase family 19 and exhibits exceptional resistance to proteolysis. The initially observed crystals, which diffracted to a resolution of 2.0 A, were improved through modification of the crystallization protocol. Well ordered crystals were subsequently obtained using N-acetyl-D-glucosamine, the monomer resulting from the hydrolysis of chitin, as an additive to the crystallization solution. Here, the characterization of a chitinase crystal that belongs to the monoclinic space group P2(1), with unit-cell parameters a = 69.08, b = 44.79, c = 76.73 A, beta = 95.33 degrees and two molecules per asymmetric unit, is reported. Diffraction data were collected to a resolution of 1.8 A. Structure refinement is currently in progress.

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References

Sali A, Blundell T . Comparative protein modelling by satisfaction of spatial restraints. J Mol Biol. 1993; 234(3):779-815. DOI: 10.1006/jmbi.1993.1626. View

Brunner F, Stintzi A, Fritig B, Legrand M . Substrate specificities of tobacco chitinases. Plant J. 1998; 14(2):225-34. DOI: 10.1046/j.1365-313x.1998.00116.x. View

Henrissat B, Bairoch A . New families in the classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem J. 1993; 293 ( Pt 3):781-8. PMC: 1134435. DOI: 10.1042/bj2930781. View

Zhu Z, Zheng T, Homer R, Kim Y, Chen N, Cohn L . Acidic mammalian chitinase in asthmatic Th2 inflammation and IL-13 pathway activation. Science. 2004; 304(5677):1678-82. DOI: 10.1126/science.1095336. View

Kezuka Y, Ohishi M, Itoh Y, Watanabe J, Mitsutomi M, Watanabe T . Structural studies of a two-domain chitinase from Streptomyces griseus HUT6037. J Mol Biol. 2006; 358(2):472-84. DOI: 10.1016/j.jmb.2006.02.013. View

Ubhayasekera W, Tang C, Ho S, Berglund G, Bergfors T, Chye M . Crystal structures of a family 19 chitinase from Brassica juncea show flexibility of binding cleft loops. FEBS J. 2007; 274(14):3695-3703. DOI: 10.1111/j.1742-4658.2007.05906.x. View

Hoell I, Dalhus B, Heggset E, Aspmo S, Eijsink V . Crystal structure and enzymatic properties of a bacterial family 19 chitinase reveal differences from plant enzymes. FEBS J. 2006; 273(21):4889-900. DOI: 10.1111/j.1742-4658.2006.05487.x. View

Gill S, von Hippel P . Calculation of protein extinction coefficients from amino acid sequence data. Anal Biochem. 1989; 182(2):319-26. DOI: 10.1016/0003-2697(89)90602-7. View

Hoell I, Klemsdal S, Vaaje-Kolstad G, Horn S, Eijsink V . Overexpression and characterization of a novel chitinase from Trichoderma atroviride strain P1. Biochim Biophys Acta. 2005; 1748(2):180-90. DOI: 10.1016/j.bbapap.2005.01.002. View

10.

Hahn M, Hennig M, Schlesier B, Hohne W . Structure of jack bean chitinase. Acta Crystallogr D Biol Crystallogr. 2000; 56(Pt 9):1096-9. DOI: 10.1107/s090744490000857x. View

11.

Hart P, Pfluger H, Monzingo A, Hollis T, Robertus J . The refined crystal structure of an endochitinase from Hordeum vulgare L. seeds at 1.8 A resolution. J Mol Biol. 1995; 248(2):402-13. View

12.

Beintema J . Structural features of plant chitinases and chitin-binding proteins. FEBS Lett. 1994; 350(2-3):159-63. DOI: 10.1016/0014-5793(94)00753-5. View

13.

Huet J, Wyckmans J, Wintjens R, Boussard P, Raussens V, Vandenbussche G . Structural characterization of two papaya chitinases, a family GH19 of glycosyl hydrolases. Cell Mol Life Sci. 2006; 63(24):3042-54. PMC: 11136075. DOI: 10.1007/s00018-006-6320-3. View

14.

Matthews B . Solvent content of protein crystals. J Mol Biol. 1968; 33(2):491-7. DOI: 10.1016/0022-2836(68)90205-2. View