Open AccessExpression, purification, crystallization and preliminary X‐ray diffraction analysis of an essential lipoprotein implicated in cell‐wall biosynthesis in Mycobacteria
Author(s) -
Marland Zara,
Beddoe Travis,
ZakerTabrizi Leyla,
Coppel Ross L.,
Crellin Paul K.,
Rossjohn Jamie
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309105037541
Subject(s) - biosynthesis , crystallization , cell wall , lipoarabinomannan , glycolipid , mycobacterium tuberculosis , crystallography , chemistry , biochemistry , microbiology and biotechnology , biology , tuberculosis , medicine , enzyme , organic chemistry , pathology
Mycobacterium tuberculosis is a renewed cause of devastation in the developing world. Critical to the success of this re‐emerging pathogen is its unusual waxy cell wall, which is rich in rare components including lipoarabinomannan (LAM) and its precursors, the phosphatidylinositol mannosides (PIMs). Balanced synthesis of these related glycolipids is intrinsic to both cell‐wall integrity and virulence in M. tuberculosis and presents a promising, albeit poorly defined, therapeutic target. Here, the expression, purification and crystallization of an essential 600‐amino‐acid lipoprotein, LpqW, implicated in this process are reported. Crystals of LpqW were grown using 20–24%( w / v ) PEG 4000, 8–16%( v / v ) 2‐propanol, 100 m M sodium citrate pH 5.5 and 10 m M DTT. A complete data set was collected at 2.4 Å using synchrotron radiation on a crystal belonging to space group C 222, with unit‐cell parameters a = 188.57, b = 312.04, c = 104.15 Å. Structure determination is under way.