title: Isolation of fusion proteins containing SecY and SecE, components of the protein translocation complex from the halophilic archaeon Haloferax volcanii.
abstract: By exploiting the salt-insensitive interaction of the cellulose-binding domain (CBD) of the Clostridium thermocellum cellulosome with cellulose, purification of CBD-fused versions of SecY and SecE, components of the translocation apparatus of the halophilic archaeon Haloferax volcanii, was undertaken. Following transformation of Haloferax volcanii cells with CBD-SecY- or -SecE-encoding plasmids, cellulose-based purification led to the capture of stably expressed, membrane-bound 68 and 25 kDa proteins, respectively. Both fusion proteins were recognized by antibodies raised against the CBD. Thus, CBD-cellulose interactions can be employed as a salt-insensitive affinity purification system for the capture of complexes containing the Haloferax volcanii translocation apparatus components SecY and SecE.
authors.affiliation: Department of Life Sciences, Ben Gurion University of the Negev, PO Box 653, Beersheva 84105, Israel.
authors.forename: Vered
authors.initials: V
authors.lastname: Irihimovitch
authors.forename: Gabriela
authors.initials: G
authors.lastname: Ring
authors.forename: Tsiona
authors.initials: T
authors.lastname: Elkayam
authors.forename: Zvia
authors.initials: Z
authors.lastname: Konrad
authors.forename: Jerry
authors.initials: J
authors.lastname: Eichler
mesh_terms: D019843:Archaeal Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D001483:Base Sequence; D002482:Cellulose; D019641:DNA, Archaeal / Q000235:genetics; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D015870:Gene Expression; D019766:Genes, Archaeal; D019622:Haloferax volcanii / Q000737:chemistry* / Q000235:genetics; D015151:Immunoblotting; D008970:Molecular Weight; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D000069816:SEC Translocation Channels
chemical_list: D019843:Archaeal Proteins; D019641:DNA, Archaeal; D029968:Escherichia coli Proteins; D011993:Recombinant Fusion Proteins; D000069816:SEC Translocation Channels; C065493:SecE protein, E coli; C057841:SecY protein, E coli; D002482:Cellulose