Orientation and nearest neighbor analysis of psbI gene product in the photosystem II reaction center complex using bifunctional cross-linkers

Abstract

Two distinct cross-linked products containing psbI gene product were generated in the photosystem II reaction center complex from spinach by treatment with bifunctional reagents directed to amino groups. The first product, which was generated by a 3,3'-dithiobis(succinimidyl propionate) treatment, is deduced to be formed between the ϵ-amino group of Lys 3 of the psbI gene product, and a side-chain amino group present on the stromal extension of the D2 protein. The CNBr cleavage analysis of the cross-linked product predicted that the amino group of the D2 protein engaged in the cross-linking is either one of the three lysine residues on the N-terminal fragment (from N-terminus to Met 19) or the Lys 268 on the 6th fragment (from Val 248 to Met 275). The second product, which was also generated on the stromal side by a 1,6-hexamethylene diisocyanate treatment preferentially in alkaline conditions, is predicted to be formed between the ϵ-amino group of Lys 3 of the psbI gene product and the N-terminal α-amino group of the α-subunit of cytochrome b 559.