Amelioration of cataracts and proteolysis in cultured lenses by cysteine protease inhibitor E64.
Cataracts were produced in cultured rat lenses by either 10 microM calcium ionophore A23187, 25 microM sodium selenite, or 30 mM xylose. E64, an inhibitor of cysteine proteases, such as calpain (EC, 3.4.22.17), reduced severity of cataract and proteolysis of crystallins when included at a 500 microM concentration in the culture medium along with cataractogenic agents. Calpain II enzyme activity and the amount of calpain antigen were decreased in the cytosol of cataractous lens. However, E64 caused an increase in the amount of an 80-kD calpain subunit associated with the ethyleneglycol-bis-(beta-aminoethylether) tetraacetic acid/ethylenediaminetetraacetic acid-washed insoluble proteins when lenses were incubated with cataractous agents. These data indicate that E64 was at least partially effective in inhibiting lens calpain, and that activation of lens calpain may involve binding to the insoluble fraction. These results provide strong evidence for the activation of calpain in rodent cataracts and suggest testing inhibitors of calpain as anticataract drugs.
Shearer
T R
TR
Department of Biochemistry, School of Dentistry, Oregon Health Sciences University, Portland 97201.
Azuma
M
M
David
L L
LL
Murachi
T
T
D000818:Animals; D000001:Calcimycin; D002154:Calpain / Q000037:antagonists & inhibitors / Q000378:metabolism; D002386:Cataract / Q000139:chemically induced / Q000201:enzymology / Q000517:prevention & control*; D002851:Chromatography, High Pressure Liquid; D003459:Crystallins / Q000378:metabolism; D046508:Culture Techniques; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D007908:Lens, Crystalline / Q000187:drug effects* / Q000201:enzymology; D007930:Leucine / Q000031:analogs & derivatives* / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D012643:Selenium; D018038:Sodium Selenite; D014994:Xylose
D003459:Crystallins; D015853:Cysteine Proteinase Inhibitors; D000001:Calcimycin; D014994:Xylose; D002154:Calpain; D007930:Leucine; D012643:Selenium; D018038:Sodium Selenite; C024974:E 64