Fluorescence resonance energy transfer mapping of subunitδ in spinach chloroplast F1 ATPase

Despite the considerable progress in the field of F0F1-ATPases caused by solving the 2.8-Angstrom structure of mitochondrial F-1 ATPase [Abrahams, J. P., Leslie, A. G. W., Lutter, R. & Walker, J. E. (1994) Nature 370, 621-628], little is known about the position and function of the enzyme's small subunits which were not resolved in the X-ray analysis. We have previously genetically engineered Cys residues into the delta subunit of chloroplast F-1 and used these mutant subunits in cross-linking studies [Lill, H., Hensel, F, Junge, W. & Engelbrecht, S. (1996) J. Biol. Chern. 271, 32737-32742]. In this work, various fluorophores have been introduced into the mutant delta subunits and used in fluorescence-resonance energy-transfer measurements. The resulting distances were fitted into the framework of existing data. Subunit delta was found to be located between two alpha/beta couples, stretching from the level of the nucleotide binding sites up to a position close to the N-termini of subunits alpha and beta. These results corroborate and further refine the previously found location of spinach CF1 delta at the periphery and membrane-distal part of CF1, where it may constitute a part of a stator in the rotatory machinery of F0F1.