Reconstitution of a light-stimulated adenylate cyclase from retina and Neurospora crassa preparations. Characterization of the heterologous systems using normal and degenerative retinas.
Adenylate cyclase catalytic subunits from Neurospora crassa membranes may interact with regulatory factors from membranes of bovine retinal rod outer segments (pretreated with N-ethylmaleimide), reconstituting a heterologous system which, in the presence of light, is catalytically active in assay mixtures containing MgATP. Maximal activation was observed at 550 nm. Transducin-depleted retinal membranes were not capable of reconstituting the heterologous light-stimulated adenylate cyclase system. Addition of a transducin preparation to depleted membranes restored the reconstitution capacity of these membranes. A similar heterologous adenylate cyclase system was reconstituted with Neurospora and mouse retinal whole membranes (pretreated with N-ethylmaleimide). Membranes from mice suffering photoreceptor degeneration (rd homozygotes) did not reconstitute an heterologous adenylate cyclase system.
Muschietti
J P
JP
Instituto de Investigaciones en Ingeniería Genética y Biología Molecular, Buenos Aires, Argentina.
Bianchini
G M
GM
Martinetto
H E
HE
Carricarte
V C
VC
Giusto
N
N
Farber
D B
DB
Torres
H N
HN
Flawia
M M
MM
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D000262:Adenylyl Cyclases / Q000032:analysis*; D000818:Animals; D002417:Cattle; D006152:Cyclic GMP / Q000378:metabolism; D008027:Light; D051379:Mice; D008810:Mice, Inbred C57BL; D009491:Neurospora / Q000201:enzymology*; D009492:Neurospora crassa / Q000201:enzymology*; D012160:Retina / Q000201:enzymology*
D000246:Adenosine Diphosphate Ribose; D000262:Adenylyl Cyclases; D006152:Cyclic GMP