PURPOSE To study the effect of storage temperature on lens crystallins quality for proteomic analysis, using αA-crystallins as internal marker. EXPERIMENTAL DESIGN Lenses were stored at -40°C, -10°C and ice for up to 10 days. Protein extracts were prepared from samples stored at -40°C and -10°C on completion of 10 days; for samples kept under ice-storage, lenses were taken out at every 24 h, ...

The ontogeny and specificity of human lens proteins were studied by immunologic and starch gel electrophoretic methods. Nine antigens were detected in noncataractous lenses by immunoelectrophoresis, while 16 components were formed on starch gel analysis. Of the different antigens, only one was specific for man and anthropoid apes. This antigen which reflected a high degree of species specificit...

BACKGROUND The eye lens is composed of fiber cells that are filled with α-, β- and γ-crystallins. The primary function of crystallins is to maintain the clarity of the lens through ordered interactions as well as through the chaperone-like function of α-crystallin. With aging, the chaperone function of α-crystallin decreases, with the concomitant accumulation of water-insoluble, light-scatterin...

Age-related cataract is a result of crystallins, the predominant lens proteins, forming light-scattering aggregates. In the low protein turnover environment of the eye lens, the crystallins are susceptible to modifications that can reduce stability, increasing the probability of unfolding and aggregation events occurring. It is hypothesized that the α-crystallin molecular chaperone system recog...

METHODS. Recombinant human C-crystallin was mixed with various concentrations of glutathione (GSH) and diamide at 25°C for 1 hour. The extent of glutathiolation of the Ccrystallin was determined by mass spectrometry. Native and S-glutathiolated C-crystallins were labeled with I, and proteolytic degradation was determined using both lens fiber lysate and reticulocyte lysate as sources of ubiquit...

Proteins exposed to UV radiation are subject to irreversible photodamage through covalent modification of tryptophans (Trps) and other UV-absorbing amino acids. Crystallins, the major protein components of the vertebrate eye lens that maintain lens transparency, are exposed to ambient UV radiation throughout life. The duplicated beta-sheet Greek key domains of beta- and gamma-crystallins in hum...

PURPOSE To determine if limited proteolysis of beta-crystallins is associated with insolubilization of proteins in rats lens during maturation and to test if the protease, calpain II, is involved. METHODS Soluble and insoluble lens proteins from 4-day-old to 4-month-old rat lens cortexes and nuclei were separated by two-dimensional electrophoresis. The insoluble proteins from 4-month-old nucl...

We have recently demonstrated that singlet oxygen-mediated photooxidation can cause an increase in blue fluorescence and covalent cross-linking in bovine lens crystallins in vitro. Because these changes closely parallel modifications known to occur in human crystallins during aging and cataractogenesis, it was suggested that singlet oxygen may play an important role in these processes in vivo. ...

PURPOSE To characterize age-related changes to proteins in the center of the human lens. METHODS Human lenses of different ages were dissected using trephines. Sucrose density gradient centrifugation was used to separate the proteins from two defined nuclear regions. Densitometry of Coomassie-stained protein bands was compared with lipid analysis with the use of mass spectrometry. RESULTS A...

A quantitative analysis of cell division and cell elongation was carried out during lens morphogenesis in the rat. At 13 days of development elongating cells in the posterior part of the lens vesicle (presumptive fibre cells) have a lower mitotic activity than cells in the anterior vesicle. By 14 days these elongating cells do not divide. Thus at 14 days of development the lens can be separated...