Transmission photoemission electron microscopy for lateral mapping of the X-ray absorption structure of a metalloprotein in a liquid cell.
We use photoemission electron microscopy in an X-ray transmission mode for full-field imaging of the X-ray absorption structure of copper in the respiratory metalloprotein hemocyanin KLH1. It contains 160 oxygen binding sites. Each site reversibly binds one molecule oxygen between two copper atoms. In our setup, hemocyanin is dissolved in aqueous solution and enclosed in an ultra-high vacuum compatible liquid sample cell with silicon nitride membranes. The local X-ray absorption structure of the liquid sample is converted into photoelectrons at the microscope side of the cell acting as a photocathode. In this way, different copper valencies are laterally distinguished under in vivo-like conditions, attributed to Cu(I) in the deoxy-state and Cu(II) in the oxy-state.
Panzer
D
D
Institut für Physik, Johannes Gutenberg-Universität, Staudinger Weg 7, Mainz, Germany.
Beck
C
C
Maul
J
J
Möller
M
M
Decker
H
H
Schönhense
G
G
D001665:Binding Sites; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006433:Hemocyanins / Q000737:chemistry* / Q000648:ultrastructure*; D046529:Microscopy, Electron, Transmission / Q000295:instrumentation* / Q000379:methods; D010100:Oxygen / Q000737:chemistry*; D011485:Protein Binding; D011487:Protein Conformation; D013048:Specimen Handling / Q000295:instrumentation* / Q000379:methods; D013052:Spectrometry, X-Ray Emission / Q000379:methods*
D006433:Hemocyanins; D010100:Oxygen