The role of positively charged residues in CXCR4 recognition probed with synthetic peptides

A high positive charge is the common characteristic shared by the beta-shee t region of stromal cell-derived factor-1 (SDF-1) and CXCR4 antagonists suc h as ALX40-4C consisting of nine D-arginines. This raises the question that the positively charged residues may play a role in recognition of CXCR4. T o test this hypothesis, two studies were carried out using synthetic peptid es. In the first study, peptide analogs possessing amino acid sequences fro m both the N-terminus and the beta-sheet region of SDF-1 were used as model s to study the functional role of the beta-sheet region of SDF-1. The attac hment of positively charged residues to the N-terminal peptide sequence of SDF-1 was found to enhance the ability of the peptides in CXCR4 binding and inhibiting CXCR4-mediated T-tropic HIV-1 entry. In the second study, two p eptides containing nine arginines and the N-terminal signal sequence of SDF -1 were used as models to study the receptor binding mechanism of CXCR4 ant agonists of high positive charges such as ALX40-4C. One peptide did not sho w signaling activity as indicated by the lack of calcium influx while anoth er peptide induced unusual calcium influx distinct from that induced by the SDF-1 N-terminal peptide. In addition, the signal induced by the SDF-1 N-t erminal peptide was inhibited by ALX40-4C. Therefore, the first study provi des experimental support for the role of the highly positive beta-sheet reg ion of SDF-1 in CXCR4 binding. The second study suggests that the binding s ite of ALX40-4C in CXCR4 may partially overlap with that of the SDF-1 N-ter minal peptide. Both findings should be valuable for the design of SDF-1 ago nists and antagonists. (C) 1999 Academic Press.