title: Highly efficient precipitation of phosphoproteins using trivalent europium, terbium, and erbium ions.
abstract: This study describes a highly efficient method for the selective precipitation of phosphoproteins by trivalent europium, terbium, and erbium metal ions. These metal cations belong to the group of lanthanides and are known to be hard acceptors with an overwhelming preference for oxygen-containing anions such as phosphates to which they form very tight ionic bonds. The method could be successfully applied to specifically precipitate phosphoproteins from complex samples including milk and egg white by forming solid metal-protein complexes. Owing to the low solubility product of the investigated lanthanide salts, the produced metal-protein complexes showed high stability. The protein pellets were extensively washed to remove nonphosphorylated proteins and contaminants. For the analysis of proteins the pellets were first dissolved in 30 % formic acid and subjected to matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) MS. For peptide mass-fingerprint analysis the precipitated phosphoproteins were enzymatically digested using microwave-assisted digestion. The method was found to be highly specific for the isolation and purification of phosphoproteins. Protein quantification was performed by colorimetric detection of total precipitated phosphoproteins and revealed more than 95 % protein recovery for each lanthanide salt.
authors.affiliation: Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens University, Innsbruck, Austria.
authors.forename: Yüksel
authors.initials: Y
authors.lastname: Güzel
authors.forename: Matthias
authors.initials: M
authors.lastname: Rainer
authors.forename: Munazza Raza
authors.initials: MR
authors.lastname: Mirza
authors.forename: Günther K
authors.initials: GK
authors.lastname: Bonn
mesh_terms: D000818:Animals; D002417:Cattle; D002645:Chickens; D004527:Egg Proteins / Q000737:chemistry / Q000302:isolation & purification*; D004529:Egg White / Q000737:chemistry; D004871:Erbium / Q000737:chemistry; D005063:Europium / Q000737:chemistry; D005590:Fractional Precipitation / Q000379:methods*; D008894:Milk Proteins / Q000737:chemistry / Q000302:isolation & purification*; D010750:Phosphoproteins / Q000737:chemistry / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013725:Terbium / Q000737:chemistry
chemical_list: D004527:Egg Proteins; D008894:Milk Proteins; D010750:Phosphoproteins; D013725:Terbium; D005063:Europium; D004871:Erbium