Expression, purification and preliminary crystallographic studies of a single-point mutant of Mos1 mariner transposase.
A soluble single-point mutant of full-length Mos1 mariner transposase (MW = 40.7 kDa) has been overexpressed in Escherichia coli, purified to 95% homogeneity and crystallized. This provides the first example of the crystallization of a eukaryotic transposase. The native crystals diffract to 2.5 A resolution and show tetragonal symmetry, with unit-cell parameters a = b = 44.5, c = 205.6 A. Multiple-wavelength anomalous data from a selenomethionyl form of the protein and data from a heavy-atom derivative have been collected.
Richardson
Julia M
JM
School of Biological Sciences, University of Edinburgh, The King's Buildings, Mayfield Road, Edinburgh EH9 3JR, Scotland. jrichard@staddmail.ed.ac.uk
Zhang
Lei
L
Marcos
Severine
S
Finnegan
David J
DJ
Harding
Marjorie M
MM
Taylor
Paul
P
Walkinshaw
Malcolm D
MD
D000595:Amino Acid Sequence; D003460:Crystallization; D018360:Crystallography, X-Ray; D029721:Drosophila Proteins / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D017354:Point Mutation; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012645:Selenomethionine; D019895:Transposases / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification
D029721:Drosophila Proteins; D011994:Recombinant Proteins; D012645:Selenomethionine; D019895:Transposases