Herbicides photoaffinity labels

Figure 4. Structural formulas of herbicidal photoaffinity labels.

Gardner, G. (1981). Azidoatrazine Photoaffinity label for the site of triazine herbicide action in chloroplasts. Science, 211 937-940. Gingrich, J.C., J.S. Buzby, V.L. Stirewalt, and D.A. Bryant (1988). Genetic analysis of two new mutations resulting in herbicide resistance in the cyanobacterium Synechococcus-sp pcc 7002. Photosyn. Res., 16 83-100. 

Pfister, K., K.E. Steinback, G. Gardner, and C.J. Amtzen (1981). Photoaffinity labeling of an herbicide receptor protein in chloroplast membranes. Proc. Natl. Acad. Sci., 78 981-985. 

Herbicides that inhibit photosynthetic electron flow prevent reduction of plastoquinone by the photosystem II acceptor complex. The properties of the photosystem II herbicide receptor proteins have been investigated by binding and displacement studies with radiolabeled herbicides. The herbicide receptor proteins have been identified with herbicide-derived photoaffinity labels. Herbicides, similar in their mode of action to 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) bind to a 34 kDa protein, whereas phenolic herbicides bind to the 43-51 kDa photosystem II reaction center proteins. At these receptor proteins, plastoquinone/herbicide interactions and plastoquinone binding sites have been studied, the latter by means of a plastoquinone-deriv-ed photoaffinity label. For the 34 kDa herbicide binding protein, whose amino acid sequence is known, herbicide and plastoquinone binding are discussed at the molecular level. 

The phenolic photoaffinity label azidodinoseb (Figure 4) binds less specifically than either azidoatrazine or azidotriazinone (14). In addition to other proteins, it labels predominantly the photosystem II reaction center proteins (spinach 43 and 47 kDa Chlamydomo-nas 47 and 51 kDa) (17). Because of the unspecific binding of azidodinoseb, this can best be seen in photosystem II preparations (17). Thus, the phenolic herbicides bind predominantly to the photosystem II reaction center, which might explain many of the differences observed between "DCMU-type" and phenolic herbicides (9). The photosystem II reaction center proteins and the 34 kDa herbicide binding protein must be located closely to and interact with each other in order to explain the mutual displacement of both types of herbicides (8,12,21). Furthermore, it should be noted that for phenolic herbicides, some effects at the donor side of photosystem II (22) and on carotenoid oxidation in the photosystem II reaction center have been found (23). 

In conclusion, observations made in the last few years, especially the binding studies with radiolabeled herbicides, the photoaffinity labeling technique, and the advances of molecular biology have substantially added to our knowledge of the mechanism of action of photosynthetic herbicides. However, many questions also remain to be answered. 

Photoaffinity labels are an efficient tool for identification of inhibitor binding proteins in the photosynthetic electron transport chain. [ H]Azido-dinoseb, an azido-deri-vative of the phenolic herbicide dinoseb, was synthesized almost a decade ago and was shown to bind primarily to a 41 kDa protein (1,2). Contrary, labeling with azido-deri-vates of diuron-type herbicides revealed that these herbicides bind to a 32 kDa protein, which has now been recognized as the D-1 protein of the photosystem II reaction center core complex (see references in (3)). Tyrosine residues in positions 237 and 254 of the D-1 sequence were demonstrated to be the primary target of [ CJazido-monuron (3). The phenolic herbicide [ I]azido-ioxynil also labels predominantly the D-1 protein in position of Val249 and only in trace amounts a 41 kDa protein (4). 

Use of 3(3-azido-4 chlorophenyl)-1,1-dimethylurea (azidomonuron), a photoaffinity labelling analogue of the herbicidal phenylurea 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), has revealed Tyr 237 and Tyr 254 of D1 to be close to the herbicide binding site (1). An interesting property of DCMU is its ability, under certain conditions, to displace a bicarbonate ion which normally binds to PS2 in the region of the non-heme ferrous atom situated between and Qg. 

The distinctly different behavior of the phenol-type herbicides following trypsin treatment suggests that different determinants within the PS II protein complex establish the "domains" that regulate the binding properties of these inhibitors. In spite of the fact that phenol-type herbicides will displace bound radiolabeled herbicides such as diuron, these inhibitors show noncompetitive inhibition (29, 30). At present, there are three lines of evidence which favor TH e involvement of two domains within the PS II complex that participate in creating the binding sites for these herbicides (a) isolated PS II particles can be selectively depleted of a polypeptide with parallel loss of atrazine sensitivity, but not dinoseb inhibition activity (33) (b) in resistant weed biotypes, chloroplast membranes that exhibit extreme triazine resistance have increased sensitivity to the phenol-type herbicides (13) and (c) experiments with azido (photoaffinity) derivatives of phenol and triazine herbicides result in the covalent labeling of different PS II polypeptides (, 31). 

---