title: Renaturation and stabilization of the telomere-binding activity of Saccharomyces Cdc13(451-693)p by L-arginine.
abstract: Production of recombinant proteins can be valuable in studying their biological functions. However, recombinant proteins expressed in Escherichia coli sometimes form undesirable insoluble aggregates. Solubilization and renaturation of these aggregates becomes a problem that one needs to solve. Here we used recombinant Cdc13(451-693)p as example to show the presence of l-arginine during renaturation greatly enhanced the renaturation efficiency. Cdc13p is the single-stranded telomere-binding protein of yeast Saccharomyces cerevisiae. The telomere-binding domain has been mapped within amino acids 451-693 of Cdc13p, Cdc13(451-693)p. Recombinant Cdc13(451-693)p was expressed in E. coli as insoluble protein aggregates. Purification of insoluble Cdc13(451-693)p was achieved by denaturing the protein with 6 M guanidine-HCl and followed by Ni-nitrilotriacetic acid agarose column chromatography. Renaturation of Cdc13(451-693)p to the active form was achieved by dialyzing denatured protein in the presence of l-arginine. Moreover, the presence of l-arginine was also helped in maintaining the telomere-binding activity of Cdc13(451-693)p. Taking together, l-arginine might have a general application in renaturation of insoluble aggregates.
authors: Institute of Biopharmaceutical Science, National Yang-Ming University, Shih-Pai, Taipei, Taiwan, 112, Republic of China.
authors: Y C
authors: YC
authors: Lin
authors: J W
authors: JW
authors: Shih
authors: C L
authors: CL
authors: Hsu
authors: J J
authors: JJ
authors: Lin
mesh_terms: D001120:Arginine / Q000494:pharmacology*; D001483:Base Sequence; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D020673:Protein Renaturation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D004268:DNA-Binding Proteins; D011994:Recombinant Proteins; D001120:Arginine