LdARL-1 His-tagged recombinant protein: purification by immobilized metal affinity expanded bed adsorption.
Previously we have cloned three ADP-ribosylation factor-like (ARL) genes from the parasitic protozoan Leishmania donovani: LdARL-3A and 3B, LdARL-1. LdARL-3A was previously purified as an active native form, which was able to bind GTP in vitro. In this paper, we have performed the production and the purification of Histidine-tagged (His-tagged) LdARL-1 recombinant protein by immobilized metal affinity chromatography (IMAC) using expanded bed adsorption (EBA) technology. This protein was purified with more than 95% purity and could be successfully used for GTP-binding assay.
Sahin
Annelise
A
CNRS UMR 5162 Génomique Fonctionnelle des Trypanosomatides, Université Victor Segalen Bordeaux 2, 146 rue Léo Saignat, 33076 Bordeaux cedex, France.
Tetaud
Emmanuel
E
Merlin
Gilles
G
Santarelli
Xavier
X
D020727:ADP-Ribosylation Factors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D006639:Histidine / Q000737:chemistry; D007893:Leishmania donovani / Q000235:genetics; D015800:Protozoan Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification
C501363:LdARL-1 His protein, Leishmania donovani; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006639:Histidine; D020727:ADP-Ribosylation Factors