Metal complexes as artificial proteases in proteomics: A palladium(II) complex cleaves various proteins in solutions containing detergents

Most popular agents for site-specific protein cleavage are proteolytic enzymes. Because they become denatured and inactivated by detergents, enzymes are inconvenient for proteomic analysis of hydrophobic proteins which require detergents as solubilizing agents. We used cis-[Pd(en)(H 2O) 2] 2+ (in which en represents ethylenediamine) as an artificial protease to effect cleavage of three bovine proteins, namely ubiquitin, β-casein, and serum albumin, in separate experiments. Cleavage took place in aqueous solutions containing 1.0% wt./vol. of either 3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) or Zwittergent 3-14 at 2.5 < pH < 2.9 and 55–60 °C for 3–72 h. Digests were separated by HPLC and analyzed by tandem mass spectrometry. Peptides were identified by de novo sequencing and matched against the bovine genome. Because cleavage by Pd(II) complexes is rather selective and therefore infrequent, 72% of the identified peptides in the digests contained more than 10 amino acid. Palladium(II) complexes hold promise as cleavage agents in proteomics studies of membrane proteins. cis-[Pd(en)(H 2O) 2] 2+ (en = ethylenediamine) effects cleavage of three bovine proteins in aqueous solutions containing 1.0% wt./vol. of either CHAPS or Zwittergent 3-14 at 2.5 < pH < 2.9 and 55–60 °C for 3–72 h. The results show Palladium(II) complexes hold promise as cleavage agents in proteomics studies of membrane proteins. [Display omitted]