Denaturation of metalloproteins with EDTA to facilitate enzymatic digestion and mass fingerprinting.
Metal ions bound to a protein often stabilize tertiary and/or quaternary structure. Consequently, the digestion of metalloproteins that precedes analysis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry is frequently incomplete. It is demonstrated that ethylenediaminetetraacetic acid (EDTA) successfully destabilizes metalloprotein structure and thereby facilitates tryptic digestion and protein identification.
Janecki
Dariusz J
DJ
Chemistry Department, Indiana University, Bloomington, 47405, USA.
Reilly
James P
JP
D000818:Animals; D024402:Carbonic Anhydrase II / Q000737:chemistry / Q000378:metabolism; D043422:Carboxypeptidases A / Q000737:chemistry / Q000378:metabolism; D002417:Cattle; D004492:Edetic Acid / Q000737:chemistry / Q000494:pharmacology*; D008667:Metalloproteins / Q000737:chemistry* / Q000378:metabolism*; D008958:Models, Molecular; D008970:Molecular Weight; D010449:Peptide Mapping / Q000379:methods*; D011489:Protein Denaturation / Q000187:drug effects; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013482:Superoxide Dismutase / Q000737:chemistry / Q000378:metabolism; D013696:Temperature; D014357:Trypsin / Q000378:metabolism*
D008667:Metalloproteins; D004492:Edetic Acid; D013482:Superoxide Dismutase; D043422:Carboxypeptidases A; D014357:Trypsin; D024402:Carbonic Anhydrase II