Isolation of nucleic acid binding proteins: an approach for isolation of cell surface, nucleic acid binding proteins.
An approach for isolation of cell surface, nucleic acid binding proteins is described. This approach relies on affinity modification of the proteins of living cells with reactive oligonucleotides bearing a haptenic group. Covalently modified proteins were isolated by hapten-specific affinity chromatography with subsequent SDS-PAGE. Isolated 68-kDa proteins responsible for the binding of oligonucleotides were MS/MS sequenced and identified as keratin K1, keratin K10, keratin K2e, and albumin.
Chelobanov
Boris P
BP
Institute of Chemical Biology and Fundamental Medicine, Siberian Division of the Russian Academy of Sciences, Novosibirsk, Russia.
Laktionov
Pavel P
PP
Kharkova
Maria V
MV
Rykova
Elena Y
EY
Vlassov
Valentin V
VV
D002294:Carcinoma, Squamous Cell / Q000473:pathology; D045744:Cell Line, Tumor; D002846:Chromatography, Affinity; D004268:DNA-Binding Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006241:Haptens / Q000737:chemistry; D006801:Humans; D053550:Keratin-10; D053557:Keratin-2; D007633:Keratins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013058:Mass Spectrometry; D008970:Molecular Weight; D018307:Neoplasms, Squamous Cell / Q000473:pathology; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D020539:Sequence Analysis, Protein; D012709:Serum Albumin / Q000302:isolation & purification / Q000378:metabolism
D004268:DNA-Binding Proteins; D006241:Haptens; C507493:KRT10 protein, human; C089241:KRT2 protein, human; D053557:Keratin-2; D009838:Oligodeoxyribonucleotides; D012709:Serum Albumin; D053550:Keratin-10; D007633:Keratins