In silico methods are useful for predicting 3D structure of binding sites when experimental information is lack. The complex interaction between γ-crystallins and small ligands is a key element in understanding the lens transparency. In spite of the high sequence similarity of γ-crystallins, different numbers of pockets were automatically identified on their molecular surfaces. γC-crystallin ha...

PURPOSE To explore the expression of the lens crystallins (αA- and βB1-crystallin) in Xenopus laevis embryonic lens development and regeneration and to analyze the order of different crystallins generated in the regenerating lens. METHODS Real Time-PCR, Immunofluorescence, and 2D-PAGE were used to analyze the expressions of αA-crystallin and βB1-crystallin, and related factors during embryoni...

PURPOSE With aging, lens crystallins undergo post-translational modifications (PTMs) and these modifications are believed to play a major role in age-related cataract development. The purpose of the present study was to determine the protein profiles of crystallins and their PTMs in the cortical and nuclear regions within an aging human lens to gain a better understanding about changes in cryst...

Two groups of patients suffering from cataract were investigated by the lymphocyte stimulation test. One group was tested before and the other after cataract surgery. Human (total) lens crystallins and bovine alpha crystallin were used as the antigens. Lymphocytes of healthy persons showed practically no positive stimulation with either antigen. Of the patients tested before operation 38% react...

The alpha-crystallins are members of the small heat shock protein family of molecular chaperones that have evolved to minimize intracellular protein aggregation; however, they are also implicated in a number of protein deposition diseases. In this study, we employed novel mass spectrometry techniques to investigate the changes in quaternary structure associated with this switch from chaperone t...

Disulfide bonding of lens crystallins contributes to the aggregation and insolubilization of these proteins that leads to cataract. A high concentration of reduced glutathione is believed to be key in preventing oxidation of crystallin sulfhydryls to form disulfide bonds. This protective role is decreased in aged lenses because of lower glutathione levels, especially in the nucleus. We recently...

PURPOSE To examine the physical properties of human lens cell membranes as a function of age. METHODS The environment of the phospholipid head groups in fiber cell membranes from human lenses, aged 22 to 83 years, was assessed with Laurdan and two-photon confocal microscopy. The effect of mild thermal stress on head group order was studied with lens pairs in which one intact lens was incubate...

The crystallins were discovered more than 100 years ago by Mörner (1893. Untersuchungen der Proteinsubstanzen in den lichtbrechenden Medien des Auges. Z. Physiol. Chem. 18, 61-106) as the main structural proteins of the vertebrate eye lens. Since that time the major mammalian crystallins referred to as alpha-, beta-, and gamma-crystallins were characterized with respect to their genetic organiz...

PURPOSE To determine the sequence of four rat beta-crystallins, confirm the sequences by mass spectrometry, and produce a two-dimensional electrophoresis (2-DE) map of soluble crystallins in young rat lens. METHODS New or additional sequences were determined for betaB1, betaB3, betaA3, and betaA4-crystallin cDNAs from Sprague-Dawley rats, and the deduced protein sequences confirmed by mass sp...

PURPOSE The aspartyl (Asp) residues 58 and 151 in αA-crystallin, and Asp36 and Asp62 in αB-crystallin in human lenses are known to be highly isomerized with aging. We investigate structural environments of these isomerizable aspartyl residues in α-crystallins of human lenses. METHODS To perform limited proteolysis experiments of purified human αA- and αB-crystallins, endoproteinase Asp-N (EC ...