title: Carnosine ameliorates lens protein turbidity formations by inhibiting calpain proteolysis and ultraviolet C-induced degradation.
abstract: Carnosine (CAR) is an endogenous peptide and present in lens, but there is little evidence for its effectiveness in calpain-induced proteolysis inhibition and its differential effects toward different wavelengths of ultraviolet (UV) irradiation. This study aimed to develop three in vitro cataract models to compare the mechanisms underlying the protective activities of CAR. Crude crystallins extracted from porcine lenses were used for antiproteolysis assays, and purified γ-crystallins were used for anti-UV assays. The turbidity in those in vitro models mimics cataract formation and was assayed by measuring optical density (OD) at 405 nm. The effectiveness of CAR on calpain-induced proteolysis was studied at 37 and 58 °C. Patterns of proteins were then analyzed by SDS-PAGE. The turbidity was reduced significantly (p<0.05) at 60 min measurements with the increased concentration of CAR (10-300 mM). SDS-PAGE showed that the decreased intensities at both ∼28 and ∼30 kDa protein bands in heat-enhanced assays were ameliorated by CAR at ≥10 mM concentrations. In UV-B studies, CAR (200, 300 mM) reduced the turbidity of γ-crystallin significantly (p<0.05) at 6 h observations. The turbidity of samples containing γ-crystallins was ameliorated while incubated with CAR (100, 300 mM) significantly (p<0.05) following 4 h of exposure to UV-C. SDS-PAGE showed that the presence of CAR reduced UV-B-induced aggregation of γ-crystallins at ∼44 kDa and resulted in less loss of γ-crystallin following UV-C exposure. The result of modeling also suggests that CAR acts as an inhibitor of calpain. In conclusion, CAR protects lens proteins more readily by inhibiting proteolysis and UV-C-induced degradation than aggregation induced by UV-B irradiation.
authors.affiliation: Institute of Biological Chemistry, Academia Sinica , Taipei 115, Taiwan.
authors.forename: Jiahn-Haur
authors.initials: JH
authors.lastname: Liao
authors.forename: I-Lin
authors.initials: IL
authors.lastname: Lin
authors.forename: Kai-Fa
authors.initials: KF
authors.lastname: Huang
authors.forename: Pei-Ting
authors.initials: PT
authors.lastname: Kuo
authors.forename: Shih-Hsiung
authors.initials: SH
authors.lastname: Wu
authors.forename: Tzu-Hua
authors.initials: TH
authors.lastname: Wu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002154:Calpain / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002336:Carnosine / Q000378:metabolism*; D002386:Cataract / Q000378:metabolism*; D003459:Crystallins / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D007908:Lens, Crystalline / Q000378:metabolism*; D008969:Molecular Sequence Data; D059748:Proteolysis / Q000528:radiation effects; D016415:Sequence Alignment; D013552:Swine; D014466:Ultraviolet Rays / Q000009:adverse effects
chemical_list: D003459:Crystallins; D006023:Glycoproteins; C039304:calpain inhibitors; D002336:Carnosine; D002154:Calpain