Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the small subunit of isopropylmalate isomerase (Rv2987c) from Mycobacterium tuberculosis.
Two C-terminally truncated variants of the small subunit of Mycobacterium tuberculosis isopropylmalate isomerase (Rv2987c; LeuD), LeuD_1-156 and LeuD_1-168, have been cloned, heterologously expressed in Escherichia coli, purified using standard chromatographic techniques and crystallized. The crystals of LeuD_1-156 belonged to the hexagonal system (space group P6(1)22 or P6(5)22) with up to four subunits in the asymmetric unit, whereas the crystals of LeuD_1-168 belonged to the monoclinic system (space group P2(1)) with two subunits in the asymmetric unit. Both crystals diffracted X-rays to beyond 2.0 A resolution and were suitable for further crystallographic analysis.
Karuppasamy
Manikandan
M
EMBL Hamburg Outstation, c/o DESY, Notkestrasse 85, D-22603 Hamburg, Germany.
Geerlof
Arie
A
Schuldt
Linda
L
Mueller-Dieckmann
Christoph
C
Weiss
Manfred S
MS
D003001:Cloning, Molecular; D003460:Crystallization; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D007535:Isomerases / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D009169:Mycobacterium tuberculosis / Q000201:enzymology* / Q000235:genetics*; D010446:Peptide Fragments / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D017510:Protein Folding; D054680:Ribosome Subunits, Small, Bacterial / Q000201:enzymology* / Q000235:genetics*; D014961:X-Ray Diffraction / Q000379:methods
D010446:Peptide Fragments; C032241:isopropylmalate isomerase; D007535:Isomerases