1H NMR relaxation investigation of herbicides interacting with photosystem II preparations fromSpinacia oleracea L.

Abstract

The interaction of three common herbicides, paraquat, acifluorfen and alachlor, with spinach chloroplast photosystem II (PS II) was investigated by measuring1H nuclear magnetic resonance spin-lattice relaxation rates, transient nuclear Overhauser effect (NOE) and NOE spectroscopy (NOESY) spectra. Binding to PS II was detected by (i) the enhancement of single-selective relaxation rates and (ii) the decrease in the optimal mixing time providing maximal cross-peak intensity in NOESY spectra. Titration of relaxation enhancements was used to calculate the dissociation constants (K d) from the bound state for paraquat (K d = 292 ± 71 μM−1) and acifluorfen (K d = 311 ± 58 μM−1). A similarK d was apparent for alachlor. Double-selective relaxation rates allowed the isolation of dipolar relaxation terms between selected proton pairs wherefrom dynamic features of the bound state were evaluated. In all cases the motional correlation time of bound herbicide (τc = 0.1−0.4 ns at 300 K) was found two orders of magnitude slower than in the free-solution state. In the case of alachlor the E and Z isomers were observed to bind differently to PS II and a change in conformation could be hypothesized.