Cloning, purification and preliminary crystallographic analysis of a putative DNA-binding membrane protein, YmfM, from Staphylococcus aureus.
The Staphylococcus aureus protein YmfM contains a helix-turn-helix motif and is thought to be a putative DNA-binding protein which is associated with the membrane through a C-terminal hydrophobic transmembrane anchor. Truncation of the protein by the removal of this C-terminal hydrophobic segment has enabled the overexpression of a soluble domain of S. aureus YmfM (DeltaYmfM) in Escherichia coli, which has been purified and subsequently crystallized. Crystals of DeltaYmfM diffract to beyond 1.0 A resolution and belong to one of the pair of enantiomorphic tetragonal space groups P4(1)2(1)2 or P4(3)2(1)2, with unit-cell parameters a = b = 45.5, c = 72.9 A and one molecule in the asymmetric unit. The crystals of DeltaYmfM have an unusually low V(M) of 1.6 A(3) Da(-1), which is one of the lowest values observed for any protein to date. A full structure determination is under way in order to provide insights into the function of this protein.
Xu
Ling
L
Krebs Institute for Biomolecular Research, Department of Molecular Biology and Biotechnology, The University of Sheffield, Sheffield S10 2TN, England.
Sedelnikova
Svetlana E
SE
Baker
Patrick J
PJ
Rice
David W
DW
D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D003001:Cloning, Molecular / Q000379:methods; D018360:Crystallography, X-Ray / Q000379:methods; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics; D057927:Hydrophobic and Hydrophilic Interactions; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D013211:Staphylococcus aureus / Q000737:chemistry* / Q000235:genetics
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D010446:Peptide Fragments; D011994:Recombinant Proteins