Cloning, expression, purification and preliminary X-ray diffraction studies of a mycobacterial protein implicated in bacterial survival in macrophages

Adam E Shahine; Phooi Y Chan; Dene Littler; Julian Vivian; Rajini Brammananth; Paul K Crellin; Ross L Coppel; Jamie Rossjohn; Travis Beddoe

doi:10.1107/S1744309113010427

Cloning, expression, purification and preliminary X-ray diffraction studies of a mycobacterial protein implicated in bacterial survival in macrophages

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2013 May 1;69(Pt 5):566-9. doi: 10.1107/S1744309113010427. Epub 2013 Apr 30.

Authors

Adam E Shahine¹, Phooi Y Chan, Dene Littler, Julian Vivian, Rajini Brammananth, Paul K Crellin, Ross L Coppel, Jamie Rossjohn, Travis Beddoe

Affiliation

¹ Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.

Abstract

Mycobacterium species have developed numerous strategies to avoid the antimycobacterial actions of macrophages, enabling them to survive within the generally inhospitable environment of the cell. The recently identified MSMEG_5817 protein from M. smegmatis is highly conserved in Mycobacterium spp. and is required for bacterial survival in macrophages. Here, the cloning, expression, purification and crystallization of MSMEG_5817 is reported. Crystals of MSMEG_5817 were grown in 1.42 M Li2SO4, 0.1 M Tris-HCl pH 7.7, 0.1 M sodium citrate tribasic dihydrate. Native and multiple-wavelength anomalous dispersion (MAD) data sets have been collected and structure determination is in progress.

Keywords: MSMEG_5817; Mycobacterium smegmatis; macrophages; selenomethionine.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacterial Proteins / chemistry
Bacterial Proteins / genetics*
Bacterial Proteins / isolation & purification
Cloning, Molecular* / methods
Gene Expression Regulation, Bacterial*
Macrophages / microbiology*
Mycobacterium smegmatis*
X-Ray Diffraction

Substances

Bacterial Proteins