Purification of proteins specifically binding human endogenous retrovirus K long terminal repeat by affinity elution chromatography.
A novel affinity elution procedure for purification of DNA-binding proteins was developed and employed to purify to near homogeneity the proteins recognizing a 21 base pair sequence within the long terminal repeat of human endogenous retroviruses K. The approach involves loading the initial protein mixture on a heparin-agarose column and elution of protein(s) of interest with a solution of double-stranded oligonucleotide containing binding sites of the protein(s). The affinity elution has several advantages over conventional DNA-affinity chromatography: (i) it is easier and faster, permitting to isolate proteins in a 1 day-one stage procedure; (ii) yield of a target protein is severalfold higher than that in DNA-affinity chromatography; (iii) it is not necessary to prepare a special affinity support for each factor to be isolated. Theaffinity elution could be a useful alternative to conventional DNA-affinity chromatography.
Trubetskoy
D O
DO
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Milklukho-Maklaya 16/10, 117997 Moscow, Russia. dmitrii@humgen.siobc.ras.ru
Zavalova
L L
LL
Akopov
S B
SB
Nikolaev
L G
LG
D001483:Base Sequence; D002846:Chromatography, Affinity / Q000379:methods*; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D020077:Endogenous Retroviruses / Q000235:genetics*; D006801:Humans; D011485:Protein Binding; D012091:Repetitive Sequences, Nucleic Acid; D013045:Species Specificity
D017931:DNA Primers