Ca(2+)- and myristoylation-dependent association of calcineurin with phosphatidylserine.
It has been proposed that N-terminal myristoylation of calcineurin B is necessary for the membrane association of calcineurin. We tested the effects of Ca(2+) and myristoylation on the binding of calcineurin B alone or heterodimeric calcineurin to phosphatidylserine or phosphatidylcholine vesicles. In the presence of excess phosphatidylserine, 50-60% of total calcineurin associated with phosphatidylserine in a Ca(2+)-sensitive manner. Calcineurin did not associate with phosphatidylcholine. Calcineurin containing both the alpha and beta catalytic subunit isoforms bound to phosphatidylserine. Calmodulin interfered with the association of calcineurin with phosphatidylserine. In the presence of Ca(2+), myristoylated calcineurin B alone did not bind to phosphatidylcholine but did bind to phosphatidylserine, although to a lesser extent than the calcineurin heterodimer. Non-myristoylated calcineurin B alone, or calcineurin containing non-myristoylated calcineurin B did not associate with phosphatidylserine in the presence of Ca(2+). These results indicate: (i) Both isoforms of calcineurin bind to phosphatidylserine. (ii) A phospholipid binding site is located on the calcineurin B subunit. (iii) Calcineurin B myristoylation is required for the Ca(2+)-sensitive binding of calcineurin to phosphatidylserine vesicles in vitro.
Perrino
B A
BA
Department of Physiology and Cell Biology, University of Nevada School of Medicine, Reno, NV 89557, USA. perrino@physio.unr.edu
Martin
B A
BA
D000818:Animals; D001665:Binding Sites / Q000502:physiology; D019703:Calcineurin / Q000737:chemistry / Q000378:metabolism*; D002118:Calcium / Q000737:chemistry / Q000378:metabolism*; D002147:Calmodulin / Q000737:chemistry / Q000378:metabolism; D020134:Catalytic Domain; D002417:Cattle; D019814:Myristic Acid / Q000737:chemistry*; D010713:Phosphatidylcholines / Q000737:chemistry; D010718:Phosphatidylserines / Q000737:chemistry / Q000378:metabolism*; D010743:Phospholipids / Q000737:chemistry; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D002147:Calmodulin; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D010743:Phospholipids; D011994:Recombinant Proteins; D019814:Myristic Acid; D019703:Calcineurin; D002118:Calcium