Zinc porphyrin as a donor for FRET in Zn(II)cytochrome c.
We demonstrate that Zn(II) porphyrin in Zn(II)cytochrome c (Zn cyt c) is a fluorescence resonance energy transfer (FRET) donor to an Alexa660 dye acceptor. The energy transfer efficiency is dependent on the distance between the two fluorophores as shown through protein denaturation studies of five Zn cyt c variants labeled with Alexa660 in different positions. The relative quantum yield, excitation and emission energies, and labeling efficiencies of this donor-acceptor pair allow for a method of analysis based on sensitized emission of the acceptor. These studies show that Zn(II) porphyrin is an effective energy donor for measurement of molecular-scale distances by FRET.
Lee
Andrea J
AJ
Department of Chemistry, University of Rochester, Rochester, New York 14627, USA.
Ensign
Amy A
AA
Krauss
Todd D
TD
Bren
Kara L
KL
D018360:Crystallography, X-Ray; D045304:Cytochromes c / Q000737:chemistry*; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry; D008665:Metalloporphyrins / Q000737:chemistry*; D008958:Models, Molecular; D011487:Protein Conformation; D017510:Protein Folding; D015032:Zinc
D005456:Fluorescent Dyes; D008665:Metalloporphyrins; C017802:zinc hematoporphyrin; D045304:Cytochromes c; D015032:Zinc