Location and quantification of metal ions in enzymes combining polyacrylamide gel electrophoresis and particle-induced X-ray emission.
A method is presented to identify and determine the relative amounts of protein-bound metal ions in situ. Proteins or their subunits are separated by SDS-PAGE, the appropriately dried gel sections are directly scanned by a collimated proton beam of 3 MeV energy, and the characteristic X-rays produced are detected. The determination of Fe content of an iron-sulfur protein (HiPiP), as well as the Fe and Ni analysis of the hydrogenase from Thiocapsa reseopersicina, have shown the feasibility of this technique.
Szökefalvi-Nagy
Z
Z
Central Research Institute for Physics, Hungarian Academy of Sciences, Budapest.
Bagyinka
C
C
Demeter
I
I
Kovács
K L
KL
Quynh
L H
LH
D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D004798:Enzymes / Q000032:analysis*; D008670:Metals / Q000032:analysis*; D011506:Proteins / Q000032:analysis; D013052:Spectrometry, X-Ray Emission / Q000379:methods*
D004798:Enzymes; D008670:Metals; D011506:Proteins