Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of DapC (Rv0858c) from Mycobacterium tuberculosis

Abstract

N-Succinyldiaminopimelate aminotransferase from Mycobacterium tuberculosis (DAP-AT; DapC; Rv0858c) has been cloned, heterologously expressed in Escherichia coli, purified using standard chromatographic techniques and crystallized in two related crystal forms. Preliminary diffraction data analysis suggests the presence of a monomer in the asymmetric unit of the tetragonal crystal form and a dimer in the asymmetric unit of the orthorhombic crystal form.

Figure 1

Crystals of Rv0858c from M. tuberculosis. (a) Clusters of crystals of Rv0858c in the orthorhombic crystal form and (b) a crystal of Rv0858c in the tetragonal crystal form.

Figure 2

Self-rotation function, κ = 180° section, based on data collected from an orthorhombic crystal of Rv0858c. The pseudo-tetragonal packing of the molecules in this crystal form is apparent from the location of the additional twofold axes in between the crystallographic axes in this section. In addition, the self-rotation function exhibits a large peak along the c direction on the κ = 90° section (not shown). The figure was produced using the program MOLREP (Collaborative Computational Project, Number 4, 1994 ▶).