Identification of a short sequence essential for actin binding by Dictyostelium ABP-120.
Tryptic digestion of ABP-120, an actin cross-linking protein from Dictyostelium discoideum, generates a ladder of peptides differing in molecular mass by 13,000 daltons, indicating a structural repeat within the molecule. A number of peptides bind actin with the smallest having a molecular mass of 17,000 daltons (T17). Our sedimentation assays also show that a peptide of 14,000 daltons does not bind actin. Using the full-length cDNA sequence (Noegel, A., Rapp, S., Lottspeich, F., Schleicher, M., and Stewart, M. (1989) J. Cell Biol. 109, 607-618) and protein sequencing techniques, we have determined that T17 begins at residue 89 while T14 begins at residue 116. Therefore we have localized 27 amino acids which are essential for actin binding activity. This region is at the end of the molecule, distal from the repetitive beta-sheet region predicted from the cDNA sequence, and displays high sequence identity with regions in the N termini of ABP/filamin, dystrophin, beta-spectrin, and alpha-actinin.
Bresnick
A R
AR
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, New York 10461.
Warren
V
V
Condeelis
J
J
D000199:Actins / Q000378:metabolism*; D000595:Amino Acid Sequence; D002352:Carrier Proteins / Q000378:metabolism*; D004023:Dictyostelium / Q000032:analysis*; D015151:Immunoblotting; D007700:Kinetics; D008840:Microfilament Proteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000378:metabolism; D011487:Protein Conformation; D012689:Sequence Homology, Nucleic Acid; D014357:Trypsin / Q000378:metabolism; D014461:Ultracentrifugation
D000199:Actins; D002352:Carrier Proteins; D008840:Microfilament Proteins; D010446:Peptide Fragments; C060680:abpC protein, Dictyostelium; D014357:Trypsin