Purification and characterization of the heterologously expressed trehalose/maltose ABC transporter complex of the hyperthermophilic archaeon Thermococcus litoralis.
We report the purification of the maltose/trehalose transporter complex MalFGK of the hyperthermophilic archaeon Thermococcus litoralis. The complex was expressed in Escherichia coli, solubilized in dodecyl maltoside and purified with the aid of a histidine tag on one of the membrane proteins. One hundred grams of cells yielded 3 mg of pure complex. The final product showed ATPase activity at 70 degrees C and was soluble at low detergent concentration. ATPase activity was not due to dissociation of the MalK subunit from the integral membrane proteins MalF and MalG but could not be further stimulated by trehalose/maltose binding protein (TMBP), be it the native protein as isolated from T. litoralis or the soluble engineered protein. The purified native TMBP was identified as a glycoprotein.
Greller
G
G
Department of Biology, University of Konstanz, Germany.
Riek
R
R
Boos
W
W
D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D019843:Archaeal Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D006868:Hydrolysis; D008320:Maltose / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D019712:Thermococcus; D014199:Trehalose / Q000378:metabolism*
D018528:ATP-Binding Cassette Transporters; D019843:Archaeal Proteins; C432664:MalF protein, Thermococcus litoralis; C432665:MalG protein, Thermococcus litoralis; C432666:MalK protein, Thermococcus litoralis; D008565:Membrane Proteins; D011994:Recombinant Proteins; D008320:Maltose; D000255:Adenosine Triphosphate; D014199:Trehalose