Studies of ligand binding to arrestin.
A striking homology is observed between the regions 70-83 and 361-374 of the sequence of bovine arrestin and the calcium-binding loops of calmodulin and troponin C. However, the predicted alpha-helices flanking the calcium-binding site in calmodulin and troponin C are not present in arrestin. Direct measurements therefore were made in order to assess whether arrestin can bind calcium. We found that arrestin does not bind Ca2+ at physiological ionic strength, as determined by equilibrium dialysis, gel filtration, and fluorescence spectroscopy. Rapid and quantitative precipitation of arrestin occurs with Tb3+. The precipitation is reversed by EDTA and blocked by Mg2+ but not by Ca2+. Prompted by several reports, we also investigated whether nucleotides bind to arrestin. Neither ATP nor GTP binds under the conditions tested. Binding of arrestin to photolyzed, phosphorylated rhodopsin also does not influence the binding of calcium or nucleotides.
Palczewski
K
K
R. S. Dow Neurological Sciences Institute of Good Samaritan Hospital and Medical Center, Portland, Oregon 97209.
Hargrave
P A
PA
D000595:Amino Acid Sequence; D000818:Animals; D000941:Antigens / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D019393:Arrestin; D002118:Calcium / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000737:chemistry; D002417:Cattle; D011232:Chemical Precipitation; D005136:Eye Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D066298:In Vitro Techniques; D008024:Ligands; D008969:Molecular Sequence Data; D009711:Nucleotides / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D012243:Rhodopsin / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013725:Terbium / Q000737:chemistry; D014785:Vision, Ocular
D000941:Antigens; D019393:Arrestin; D002135:Calcium-Binding Proteins; D005136:Eye Proteins; D008024:Ligands; D009711:Nucleotides; D010750:Phosphoproteins; D013725:Terbium; D012243:Rhodopsin; D002118:Calcium