Chlorophyll binding to peptide maquettes containing a retention motif.

The motif Glu-X-X-His/Asn-X-Arg is conserved in the first and third membrane-spanning domains of all light-harvesting chlorophyll a/b- and a/c-binding proteins in chloroplasts. Molecular modeling of synthetic peptides containing the sequence Glu-Ile-Val-His-Ser-Arg, a motif found in the apoprotein of the major light-harvesting complex in plants, generated a loop structure formed by intrapeptide, electrostatic attraction between Glu and Arg. His, Asn, and charge-compensated Glu-Arg pairs are known ligands of the magnesium atom in chlorophyll. The prediction that this structure should bind two molecules of chlorophyll was confirmed experimentally with an assay based on fluorescence resonance energy transfer between peptides and chlorophyll a. Motifs with both potential ligands bound approximately two times the amount of chlorophyll as one in which His was replaced by Ala. These results support the conclusion that formation of this intermediate, within membranes of the envelope, is a crucial step in assembly of light-harvesting complexes and a mechanism that regulates import of the apoproteins into the chloroplast.