Molecular cloning and biochemical characterization of bovine retina calcineurin.
Calcineurin (CaN) is a member of ser/thr protein phosphatase family. Earlier, we have reported that CaN is present in all eye tissues, although the activity and protein expression varied (Seitz et al., Invest Opthalmol Vis Sci, 43:15-21, 2002). We have isolated a full-length cDNA encoding bovine retina CaN. The CaN A subunit consists of 511 amino acid residues. A 10 amino acid (ATVEAIEADE) deletion before the autoinhibitory domain was observed in bovine retina CaN A compared to bovine brain CaN A. The study on CaN activity and regulation demonstrated that different metal ions have different effects on its phosphatase activity. Ni(2+) was found to be the strongest stimulator, while Zn(2+) was found to inhibit CaN phosphatase activity. Mn(2+) was a relatively less effective stimulator compared to Ni(2+). Fe(2+) was also able to stimulate CaN phosphatase activity; in contrast, a previous study found Fe(2+) slightly inhibited CaN activity from bovine brain. The residues at 97-201 were found to be essential for bovine retina CaN A phosphatase activity. The residues at 407-456 also had an inhibitory effect on CaN A phosphatase activity in addition to the previously known autoinhibitory domain at 457-480. These observations suggest that bovine retina CaN A might possess some distinct structural characteristics.
Zuo
Yuan
Y
Department of Pathology and Laboratory Medicine College of Medicine, University of Saskatchewan and Health Research Division, Saskatchewan Cancer Agency, 20 Campus Drive, Saskatoon, SK, S7N 4H4, Canada.
Selvakumar
Ponniah
P
Sharma
Rajendra K
RK
D000595:Amino Acid Sequence; D000818:Animals; D019703:Calcineurin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002417:Cattle; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005136:Eye Proteins; D008670:Metals; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D017434:Protein Structure, Tertiary; D012160:Retina / Q000737:chemistry*
D018076:DNA, Complementary; D005136:Eye Proteins; D008670:Metals; D019703:Calcineurin; D010744:Phosphoric Monoester Hydrolases