Use of laser flash photolysis time-resolved spectrophotometry to investigate interprotein and intraprotein electron transfer mechanisms.
A description is given of the methodology developed in our laboratory for the application of laser flash photolysis to the elucidation of the kinetics and mechanism of electron transfer processes which occur intermolecularly between two protein molecules within a collisional complex, or intramolecularly between two redox centers within a single multisubunit or multidomain protein. This involves the use of flavin analogs, excited to their lowest triplet state by a laser flash, to initiate electron transfer, either by oxidation of a sacrificial donor followed by redox protein reduction via the flavin semiquinone, or by direct oxidation of a reduced redox protein by the flavin triplet. Time-resolved spectrophotometry is used to follow the course of the sequence of electron transfer events initiated by the laser flash. The application of this methodology to the following systems is described: cytochrome c/cytochrome c peroxidase; ferredoxin/ferredoxin NADP+ reductase; cytochrome c/plastocyanin; flavocytochrome b2; and sulfite oxidase.
Tollin
G
G
Department of Biochemistry, University of Arizona, Tucson 85721.
Hurley
J K
JK
Hazzard
J T
JT
Meyer
T E
TE
D000818:Animals; D002645:Chickens; D003574:Cytochrome c Group / Q000737:chemistry / Q000378:metabolism; D003578:Cytochrome-c Peroxidase / Q000737:chemistry / Q000378:metabolism; D004579:Electron Transport; D005287:Ferredoxin-NADP Reductase / Q000737:chemistry / Q000378:metabolism; D005288:Ferredoxins / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000737:chemistry / Q000378:metabolism; D042844:L-Lactate Dehydrogenase (Cytochrome); D007834:Lasers; D010084:Oxidation-Reduction; D050862:Oxidoreductases Acting on Sulfur Group Donors / Q000737:chemistry / Q000378:metabolism; D010782:Photolysis; D010970:Plastocyanin / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*; D013053:Spectrophotometry / Q000379:methods
D003574:Cytochrome c Group; D005288:Ferredoxins; D011506:Proteins; D010970:Plastocyanin; D007770:L-Lactate Dehydrogenase; D042844:L-Lactate Dehydrogenase (Cytochrome); D003578:Cytochrome-c Peroxidase; D005287:Ferredoxin-NADP Reductase; D050862:Oxidoreductases Acting on Sulfur Group Donors