title: Age-related decline in ubiquitin conjugation in response to oxidative stress in the lens.
abstract: Accumulation of damaged proteins is a major age-related change in lenses of virtually all species and is associated with lens opacification. Proteolytic removal of the damaged proteins may play an important role in maintaining the transparency of the lens. In many tissues, selective removal of abnormal or damaged proteins occurs via a ubiquitin-dependent proteolytic pathway. Ubiquitin, an 8.5 kDa polypeptide, selectively binds to proteins to form ubiquitin-protein conjugates. This ubiquitin-protein conjugate is, in most cases, a signal for protein degradation. In this work, age-related changes in rat lens in the following aspects were detected: (a) levels of the ubiquitin-protein conjugates, (b) some of the enzymes involved in ubiquitin conjugation in rat lenses, and (c) ability to respond to oxidative damage. Endogenous ubiquitin-protein conjugates were detected in epithelium, cortex and nucleus of lenses from young and old rats. The levels of endogenous high molecular weight (HMW) ubiquitin-protein conjugates in each developmental zone of the lenses from young rats were higher than that in the counterparts of lenses from old animals. Peroxide-treatment generally resulted in elevated levels of endogenous HMW ubiquitin-protein conjugates although masses of bulk proteins remain unchanged. The increases in ubiquitin-protein conjugates in the epithelial sections of young and old lenses upon oxidative stress were comparable. In the cortex of young lenses, there was a significant oxidation-related increase in ubiquitin-protein conjugates. There was a similar trend but diminished response in the cortex of old lenses. Nuclear fibers from young lenses also showed an oxidation-induced increase in the level of ubiquitin-protein conjugates. This response was not observed in nuclear fibers of old lenses. The ability to form HMW-ubiquitin conjugates with exogenous 125I-labeled ubiquitin in the lens also increased upon oxidative stress. The extent of the increase in the de-novo ubiquitin conjugating activity upon exposure to oxidation in old lens was much smaller than in young lens. Ubiquitin-activating enzyme (E1), and ubiquitin conjugating enzymes (E2(17k), E2(20k) and E2(25k) were detected by thiol ester assays or Western blot analysis. No significant age-related changes in the levels of E1, E2(17k), E2(20k) and E2(25k) were detected. The activity of E1 and E2(17k) increased upon exposure to H2O2. These data indicate that lens has the ability to increase ubiquitin conjugation activity in response to oxidative stress and this ability is attenuated upon aging. The age-related decrease in the ability to mount a ubiquitin-dependent response upon oxidation may contribute to the accumulation of damaged proteins in the old lenses.
authors.affiliation: Laboratory for Nutrition and Vision Research, Jean Mayer USDA Human Nutrition Research Center on Aging, Tufts University, MA 02111, USA.
authors.forename: F
authors.initials: F
authors.lastname: Shang
authors.forename: X
authors.initials: X
authors.lastname: Gong
authors.forename: H J
authors.initials: HJ
authors.lastname: Palmer
authors.forename: T R
authors.initials: TR
authors.lastname: Nowell
authors.forename: A
authors.initials: A
authors.lastname: Taylor
mesh_terms: D000375:Aging; D000818:Animals; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology; D066298:In Vitro Techniques; D007904:Lens Cortex, Crystalline / Q000187:drug effects / Q000378:metabolism; D007907:Lens Nucleus, Crystalline / Q000187:drug effects / Q000378:metabolism; D007908:Lens, Crystalline / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D008025:Ligases / Q000302:isolation & purification / Q000502:physiology; D018384:Oxidative Stress; D011506:Proteins / Q000378:metabolism; D051381:Rats; D044764:Ubiquitin-Activating Enzymes; D044763:Ubiquitin-Conjugating Enzymes; D044767:Ubiquitin-Protein Ligases; D014452:Ubiquitins / Q000378:metabolism*
chemical_list: D011506:Proteins; D014452:Ubiquitins; D006861:Hydrogen Peroxide; D044763:Ubiquitin-Conjugating Enzymes; D044767:Ubiquitin-Protein Ligases; D008025:Ligases; D044764:Ubiquitin-Activating Enzymes