Immunoaffinity chromatography of diadenosine 5',5'''-P1,P4-tetraphosphate phosphorylase from Saccharomyces cerevisiae.
Diadenosine 5',5'''-P1,P4-tetraphosphate (Ap4A) phosphorylase has been isolated previously using classical protein isolation techniques [A. Guranowski and S. Blanquet (1985) J. Biol. Chem. 260, 3542-3547]. A protein A-Sepharose immunoaffinity column was prepared to simplify the purification procedure. The immunoaffinity column was prepared using specific polyclonal antibodies to Ap4A phosphorylase covalently coupled to protein A-Sepharose with dimethyl pimelimidate by a modification of the procedure of C. Schneider et al. [(1982) J. Biol. Chem. 257, 10,766-10,769]. The specific activity of the immunoaffinity-purified enzyme showed an increase equivalent to the specific activity obtained by chromatography on DEAE-cellulose and hydroxyapatite columns.
Avila
D M
DM
Department of Biochemistry, University of Texas Health Science Center, San Antonio 78284-7760.
Kaushal
V
V
Barnes
L D
LD
D001709:Biotechnology; D002846:Chromatography, Affinity / Q000379:methods*; D005069:Evaluation Studies as Topic; D009713:Nucleotidyltransferases / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D012685:Sepharose / Q000031:analogs & derivatives; D013205:Staphylococcal Protein A
D009713:Nucleotidyltransferases; D012685:Sepharose; C042030:Staphylococcal protein A-sepharose; D013205:Staphylococcal Protein A; C044740:diadenosine 5,5'''-P(1),P(4)-tetraphosphate alpha,beta-phosphorylase