Overexpression and purification of elongation factor 3 from Saccharomyces cerevisiae.
The translational elongation factor 3 (EF-3) from Saccharomyces cerevisiae was overexpressed and purified to near homogeneity from the post-ribosomal supernatant fraction (S-100). A detailed protocol for the isolation of overexpressed EF-3 is presented. The procedure involves ion-exchange chromatography on DEAE-Sepharose and CM-Sepharose and affinity chromatography on ATP-agarose. A protein purity of > or = 96% was established by quantitating the silver-stained SDS/polyacrylamide gel. The present method facilitates isolation of EF-3 in large amounts in high yield.
Kambampati
R
R
Medical College of Wisconsin, Department of Biochemistry, Milwaukee 53226, USA.
Chakraburtty
K
K
D000255:Adenosine Triphosphate / Q000378:metabolism; D002455:Cell Division; D002474:Cell-Free System; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D005656:Fungal Proteins; D006868:Hydrolysis; D010445:Peptide Elongation Factors / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification*; D010455:Peptides / Q000138:chemical synthesis; D010957:Plasmids; D012270:Ribosomes / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins
D005656:Fungal Proteins; D010445:Peptide Elongation Factors; D010455:Peptides; D029701:Saccharomyces cerevisiae Proteins; C111584:YEF3 protein, S cerevisiae; C012001:polyphenylalanine; D000255:Adenosine Triphosphate