CRYSTALLIZATION OF HEVAMINE, AN ENZYME WITH LYSOZYME CHITINASE ACTIVITY FROM HEVEA-BRASILIENSIS LATEX

Hevamine, an enzyme with both lysozyme and chitinase activity, was isolated and purified from Hevea brasiliensis (rubber tree) latex. The enzyme (molecular weight 29,000) is homologous to certain "pathogenesis-related" proteins from plants, but not to hen egg-white or phage T4 lysozyme. To investigate the atomic details of the substrate specificity and the cause for hevamine's low pH optimum (pH 4.0), we have crystallized two hevamine isozymes as a first step towards a high-resolution X-ray structure determination. Suitable crystals were obtained at room temperature from hanging drop experiments by vapor diffusion against 1.7 m to 3.4 m-NaCl (pH 5.0 to 9.0) for the major isozyme, and by vapor diffusion against 2.5 m to 4.3 m-NaCl (pH 5.0 to 8.0) for the minor one. Both isozymes give the same crystal morphology and space group. Their space group is P212121 with cell dimensions a = 82.3 A ̊, b = 58.1 A ̊ and c = 52.5 A ̊ (1 Å = 0.1 nm). The crystals diffract to at least 2.0 Å resolution. © 1990.