High-level bacterial expression and purification of apicomplexan micronemal proteins for structural studies.
The cysteine-rich N-terminal domain of the micronemal adhesive protein MIC1 (MIC1-NT) from Toxoplasma gondii was cloned, expressed in Escherichia coli and purified. MIC1-NT is amenable to structural studies as shown by preliminary NMR and X-ray analysis. Positive results with two further micronemal proteins indicate that our strategy has wider application.
Saouros
S
S
Department of Biological Sciences, Division of Molecular Biosciences, Centre for Structural Biology, Imperial College London, South Kensington, London SW7 2AZ, UK.
Blumenschein
T M A
TM
Sawmynaden
K
K
Marchant
J
J
Koutroukides
T
T
Liu
B
B
Simpson
P
P
Carpenter
E P
EP
Matthews
S J
SJ
D000818:Animals; D015815:Cell Adhesion Molecules / Q000096:biosynthesis / Q000302:isolation & purification*; D002850:Chromatography, Gel; D003001:Cloning, Molecular / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D015800:Protozoan Proteins / Q000096:biosynthesis / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014122:Toxoplasma / Q000737:chemistry*
D015815:Cell Adhesion Molecules; C105877:MIC1 protein, Toxoplasma gondii; D015800:Protozoan Proteins