Open AccessOverexpression, purification and crystallization of lysine ɛ‐aminotransferase (Rv3290c) from Mycobacterium tuberculosis H37Rv
Author(s) -
Tripathi Sarvind Mani,
Ramachandran Ravishankar
Publication year - 2006
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309106016824
Subject(s) - cofactor , lysine , pyridoxal phosphate , crystallization , enzyme , pyridoxal , crystallography , chemistry , monomer , ammonium , recombinant dna , mycobacterium tuberculosis , ammonium sulfate , protein subunit , biochemistry , amino acid , chromatography , tuberculosis , organic chemistry , medicine , pathology , gene , polymer
Lysine ɛ‐aminotransferase (LAT) is a protein involved in lysine catabolism; it belongs to the aminotransferase family of enzymes, which use pyridoxal 5′‐phosphate (PLP) as a cofactor. LAT probably plays a significant role during the persistent/latent phase of Mycobacterium tuberculosis , as observed by its up‐regulation by ∼40‐fold during this stage. Crystals of recombinant LAT have been grown in 0.1 M trisodium citrate dihydrate solution containing 0.2 M ammonium acetate and 25% PEG 4000 in the pH range 5.4–6.0. Diffraction data extending to 1.98 Å were collected at room temperature from a single crystal. Crystals are trigonal in shape and belong to space group P 3 1 21, with unit‐cell parameters a = 103.26, b = 103.26, c = 98.22 Å. The crystals contain a monomer in the asymmetric unit, which corresponds to a Matthews coefficient ( V M ) of 3.1 Å 3 Da −1 .