Photoinduced reduction of the medial FeS center in the hydrogenase small subunit HupS from Nostoc punctiforme.
The small subunit from the NiFe uptake hydrogenase, HupSL, in the cyanobacterium Nostoc punctiforme ATCC 29133, has been isolated in the absence of the large subunit (P. Raleiras, P. Kellers, P. Lindblad, S. Styring, A. Magnuson, J. Biol. Chem. 288 (2013) 18,345-18,352). Here, we have used flash photolysis to reduce the iron-sulfur clusters in the isolated small subunit, HupS. We used ascorbate as electron donor to the photogenerated excited state of Ru(II)-trisbipyridine (Ru(bpy)3), to generate Ru(I)(bpy)3 as reducing agent. Our results show that the isolated small subunit can be reduced by the Ru(I)(bpy)3 generated through flash photolysis.
Department of Chemistry, Ångström Laboratory, Uppsala University, Box 523, SE-75120 Uppsala, Sweden.
Patrícia
P
Raleiras
Department of Chemistry, Ångström Laboratory, Uppsala University, Box 523, SE-75120 Uppsala, Sweden.
Leif
L
Hammarström
Department of Chemistry, Ångström Laboratory, Uppsala University, Box 523, SE-75120 Uppsala, Sweden.
Peter
P
Lindblad
Department of Chemistry, Ångström Laboratory, Uppsala University, Box 523, SE-75120 Uppsala, Sweden. Electronic address: stenbjorn.styring@kemi.uu.
Stenbjörn
S
Styring
Department of Chemistry, Ångström Laboratory, Uppsala University, Box 523, SE-75120 Uppsala, Sweden. Electronic address: ann.magnuson@kemi.uu.se.
Ann
A
Magnuson
D015082:2,2'-Dipyridyl / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D001205:Ascorbic Acid / Q000737:chemistry / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D056831:Coordination Complexes; D004578:Electron Spin Resonance Spectroscopy; D006864:Hydrogenase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007506:Iron-Sulfur Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D008956:Models, Chemical; D015394:Molecular Structure; D046937:Nostoc / Q000201:enzymology* / Q000235:genetics; D010084:Oxidation-Reduction / Q000187:drug effects / Q000528:radiation effects; D010782:Photolysis / Q000187:drug effects / Q000528:radiation effects; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D013053:Spectrophotometry; D013379:Substrate Specificity; D017975:Ruthenium Compounds
D015082:2,2'-Dipyridyl; D001205:Ascorbic Acid; D001426:Bacterial Proteins; D056831:Coordination Complexes; D006864:Hydrogenase; D007506:Iron-Sulfur Proteins; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; C547232:tris(2,2-bipyridine)-ruthenium(II); D017975:Ruthenium Compounds; C043296:nickel-iron hydrogenase
Electron paramagnetic resonance; Electron transfer; Flash photolysis; Hydrogenase; Iron–sulfur