title: Isolation of a Drosophila gene coding for a protein containing a novel phosphatidylserine-binding motif.
abstract: To elucidate the molecular basis of the binding of proteins to the membrane phospholipid phosphatidylserine (PS), we characterized PS-binding peptides isolated from a phage display library. Amino acid sequences deduced from the nucleotide sequences of over 60 phage clones isolated revealed that there was no common primary structure among these peptides, but all peptides were rich in basic amino acid residues. In particular, 15 clones encoded peptides that contained contiguous arginine residues. Characterization of two such peptides in more detail showed that they bound to PS, and to a much lower extent to other phospholipids, including phosphatidylinositol, phosphatidylethanolamine, and phosphatidylcholine. Unlike other Ca2+-dependent PS-binding proteins, these peptides did not require Ca2+ for binding to PS, and the addition of Ca2+ did not alter the phospholipid specificity. Substitution of one of the two RR sequences in one peptide by alanine had no effect, but that of both sequences completely abolished the activity. Furthermore, we identified a Drosophila gene coding for a presumed nuclear protein that shares an amino acid sequence, including a RR residue, with one of the two PS-binding peptides. This protein bound to PS partly depending on the presence of the RR residue. These results allowed us to conclude that an amino acid sequence including contiguous arginine residues is a novel motif that defines Ca2+-independent PS-binding activity.
authors.affiliation: Graduate School of Natural Science and Technology, Kanazawa University, Shizenken, Kakuma-machi, Kanazawa, Ishikawa 920-1192.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Nakai
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Nomura
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Sato
authors.forename: Akiko
authors.initials: A
authors.lastname: Shiratsuchi
authors.forename: Yoshinobu
authors.initials: Y
authors.lastname: Nakanishi
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D004330:Drosophila / Q000235:genetics*; D029721:Drosophila Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D017344:Genes, Insect; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D010718:Phosphatidylserines / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects
chemical_list: D029721:Drosophila Proteins; D019151:Peptide Library; D010455:Peptides; D010718:Phosphatidylserines; D002118:Calcium