title: Lens crystallin modifications and cataract in transgenic mice overexpressing acylpeptide hydrolase.
abstract: The accumulation of crystallin fragments in vivo and their subsequent interaction with crystallins are responsible, in part, for protein aggregation in cataracts. Transgenic mice overexpressing acylpeptide hydrolase (APH) specifically in the lens were prepared to test the role of protease in the generation and accumulation of peptides. Cataract development was seen at various postnatal days in the majority of mice expressing active APH (wt-APH). Cataract onset and severity of the cataracts correlated with the APH protein levels. Lens opacity occurred when APH protein levels were >2.6% of the total lens protein and the specific activity, assayed using Ac-Ala-p-nitroanilide substrate, was >1 unit. Transgenic mice carrying inactive APH (mt-APH) did not develop cataract. Cataract development also correlated with N-terminal cleavage of the APH to generate a 57-kDa protein, along with an increased accumulation of low molecular weight (LMW) peptides, similar to those found in aging human and cataract lenses. Nontransgenic mouse lens proteins incubated with purified wt-APH in vitro resulted in a >20% increase in LMW peptides. Crystallin modifications and cleavage were quite dramatic in transgenic mouse lenses with mature cataract. Affected lenses showed capsule rupture at the posterior pole, with expulsion of the lens nucleus and degenerating fiber cells. Our study suggests that the cleaved APH fragment might exert catalytic activity against crystallins, resulting in the accumulation of distinct LMW peptides that promote protein aggregation in lenses expressing wt-APH. The APH transgenic model we developed will enable in vivo testing of the roles of crystallin fragments in protein aggregation.
authors.affiliation: From the Departments of Ophthalmology and.
authors.forename: Puttur
authors.initials: P
authors.lastname: Santhoshkumar
authors.forename: Leike
authors.initials: L
authors.lastname: Xie
authors.forename: Murugesan
authors.initials: M
authors.lastname: Raju
authors.forename: Lixing
authors.initials: L
authors.lastname: Reneker
authors.forename: K Krishna
authors.initials: KK
authors.lastname: Sharma
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002386:Cataract / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D003459:Crystallins / Q000737:chemistry / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D006868:Hydrolysis; D007908:Lens, Crystalline / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D008822:Mice, Transgenic; D009154:Mutation; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D010447:Peptide Hydrolases / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics
chemical_list: D003459:Crystallins; D010446:Peptide Fragments; D010447:Peptide Hydrolases; C038220:acylaminoacyl-peptidase
keywords: Acylpeptide Hydrolase; Cataract; Crystallins; Lens; Peptides; Protease; Protein Degradation; Transgenic Mice