Overexpression, purification and crystallization of lysine epsilon-aminotransferase (Rv3290c) from Mycobacterium tuberculosis H37Rv.
Lysine epsilon-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 approximately 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 A were collected at room temperature from a single crystal. Crystals are trigonal in shape and belong to space group P3(1)21, with unit-cell parameters a = 103.26, b = 103.26, c = 98.22 A. The crystals contain a monomer in the asymmetric unit, which corresponds to a Matthews coefficient (V(M)) of 3.1 A3 Da(-1).
Tripathi
Sarvind Mani
SM
Molecular and Structural Biology Division, Central Drug Research Institute, PO Box 173, Chattar Manzil, Mahatma Gandhi Marg, Lucknow 226001, India.
Ramachandran
Ravishankar
R
D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification; D003460:Crystallization; D051306:L-Lysine 6-Transaminase / Q000737:chemistry* / Q000302:isolation & purification; D009169:Mycobacterium tuberculosis / Q000201:enzymology*; D011994:Recombinant Proteins; D012997:Solvents; D014961:X-Ray Diffraction
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D012997:Solvents; D051306:L-Lysine 6-Transaminase