title: Age-dependent inhibition of proteasome chymotrypsin-like activity in the retina.
abstract: The proteasome plays a fundamental role in processes essential for cell viability. A loss in proteasome function has been associated with aging, as well as a number of age-related diseases. Defining the mechanism(s) behind this loss in function will add important information regarding the molecular basis for aging. In the current study, we performed an age-based comparison of proteasome function and composition of subunits and regulatory proteins in the neural retina and retinal pigment epithelium (RPE) in Fischer 344 rats. In the RPE, there was no age-dependent difference in activity, subunit composition, or content of proteasome regulators, PA28 and PA700. In contrast, the aged neural retina demonstrated a significant reduction in the chymotrypsin-like activity and decreased degradation of both casein and casein modified by 4-hydroxynonenal. This loss in function could not be explained by differences in subunit composition, content of PA28 and PA700, or reversible modification of cysteine residues. To begin investigating the molecular basis for the age-associated decrement in proteasome function, we modified the cysteine residues in proteasome from young rats with the sulfhydryl-reactive chemical N-ethylmaleimide. We observed inhibition of the chymotrypsin-like activity and decreased degradation of casein that was comparable to that seen in aged retinas. Thus, chemical modification of cysteine provides an in vitro method that partially recapitulates aging proteasome. Further studies are required to confirm irreversible modification of functionally significant cysteine as a potential mechanism behind the age-related loss in proteasome function.
authors.affiliation: Department of Ophthalmology, 380 Lions Research Building, 2001 6th Street SE, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Rebecca J
authors.initials: RJ
authors.lastname: Kapphahn
authors.forename: Erin J
authors.initials: EJ
authors.lastname: Bigelow
authors.forename: Deborah A
authors.initials: DA
authors.lastname: Ferrington
mesh_terms: D000375:Aging / Q000378:metabolism*; D000447:Aldehydes / Q000494:pharmacology; D000818:Animals; D002364:Caseins / Q000378:metabolism; D002614:Chelating Agents / Q000378:metabolism; D002918:Chymotrypsin / Q000378:metabolism*; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005033:Ethylmaleimide / Q000494:pharmacology; D005136:Eye Proteins / Q000032:analysis / Q000378:metabolism*; D018841:HSP90 Heat-Shock Proteins / Q000032:analysis; D010857:Pigment Epithelium of Eye / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex / Q000032:analysis / Q000378:metabolism*; D061988:Proteasome Inhibitors; D011489:Protein Denaturation / Q000502:physiology; D051381:Rats; D011916:Rats, Inbred F344; D012160:Retina / Q000378:metabolism*
chemical_list: D000447:Aldehydes; D002364:Caseins; D002614:Chelating Agents; D003432:Cross-Linking Reagents; D004791:Enzyme Inhibitors; D005136:Eye Proteins; D018841:HSP90 Heat-Shock Proteins; C506159:PA28 protein, rat; C085834:PA700 proteasome activator; D061988:Proteasome Inhibitors; D002918:Chymotrypsin; D046988:Proteasome Endopeptidase Complex; C027576:4-hydroxy-2-nonenal; D005033:Ethylmaleimide