title: Purification of prostatic acid phosphatase (PAP) for structural and functional studies.
abstract: High-scale purification methods are required for several protein studies such as crystallography, mass spectrometry, circular dichroism, and function. Here we describe a purification method for PAP based on anion exchange, L-(+)-tartrate affinity, and gel filtration chromatographies. Acid phosphatase activity and protein concentration were measured for each purification step, and to collect the fractions with the highest acid phosphatase activity the p-nitrophenyl phosphate method was used. The purified protein obtained by the procedure described here was used for the determination of the first reported three-dimensional structure of prostatic acid phosphatase.
authors.affiliation: Department of Clinical Chemistry, Institute of Clinical Medicine, University of Helsinki, Helsinki, Finland.
authors.forename: Annakaisa M
authors.initials: AM
authors.lastname: Herrala
authors.forename: Ileana B
authors.initials: IB
authors.lastname: Quintero
authors.forename: Pirkko T
authors.initials: PT
authors.lastname: Vihko
mesh_terms: D000135:Acid Phosphatase; D002846:Chromatography, Affinity / Q000379:methods*; D002850:Chromatography, Gel / Q000379:methods*; D002852:Chromatography, Ion Exchange; D006801:Humans; D008297:Male; D009596:Nitrophenols / Q000737:chemistry; D009943:Organophosphorus Compounds / Q000737:chemistry; D011467:Prostate / Q000201:enzymology*; D017027:Protein Tyrosine Phosphatases / Q000737:chemistry / Q000302:isolation & purification*; D013379:Substrate Specificity; D013644:Tartrates / Q000378:metabolism
chemical_list: D009596:Nitrophenols; D009943:Organophosphorus Compounds; D013644:Tartrates; C008644:nitrophenylphosphate; D000135:Acid Phosphatase; C428439:prostatic acid phosphatase; D017027:Protein Tyrosine Phosphatases; C029768:tartaric acid