Invertebrate rhodopsin cleavage by an endogenous calcium activated protease.
Calcium-activated proteases have been purified from a number of vertebrate tissues, including the retina and lens. These proteases exhibit similar characteristics and are believed to be involved in the regulation of cytoskeletal elements. Here we report the partial purification and characterization of a calcium-activated protease from the squid photoreceptor cell which, when activated, specifically removes 10 kDa from the carboxyl-terminal of squid rhodopsin. No other detectable soluble proteins from the invertebrate photoreceptor are susceptible to cleavage and only one non-opsin, integral membrane protein shows evidence of cleavage. The enzyme requires 5 mM calcium for half maximal activation, and is not significantly activated by other divalent ions. The protease has a molecular weight of approximately 350 kDa, as determined by gel filtration, and when partially purified by casein affinity chromatography, it runs as three main bands of 76, 63 and 36 kDa on SDS-PAGE. The crude protease loses as much as 80% of its activity in 24 hr, whereas the partially purified protease is stable up to 4 weeks at 4 degrees C.
Oldenburg
K R
KR
Jules Stein Eye Institute, University of California, Los Angeles 90024-7008.
Hubbell
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WL
D000595:Amino Acid Sequence; D000818:Animals; D002154:Calpain / Q000302:isolation & purification* / Q000502:physiology; D002846:Chromatography, Affinity; D049832:Decapodiformes / Q000201:enzymology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012243:Rhodopsin / Q000378:metabolism*; D012374:Rod Cell Outer Segment / Q000201:enzymology
D012243:Rhodopsin; D002154:Calpain