PURPOSE To identify mutations in 12 genes in Chinese families with congenital cataracts. METHODS Twenty five families with congenital cataracts involved in this study. The coding exons and adjacent intronic regions of 12 genes were analyzed by cycle sequencing, including the alpha A crystallin (CRYAA), alpha B crystallin (CRYAB), beta A1 crystallin (CRYBA1), beta A4 crystallin (CRYBA4), beta ...

The mechanism of aggregation and insolubilization of lens proteins was examined based on the kinetics of crystallin-crystallin interaction determined by the surface plasmon resonance method on a BlAcore system. Lens proteins are composed mainly of three types crystallins, alpha-, beta-, and gamma-crystallin. The present study indicated that alpha-crystallin shows marked self-interaction. Furthe...

Alpha, beta, and gamma crystallins from rabbit eye lens have been prepared by continuousflow paper electrophoresis and gel filtration. These methods yielded well-defined fractions in a highly reproducible manner with essentially quantitative recovery of material. The behavior of each of the crystallins in diethylaminoethyl cellulose has been examined. Sulfhydryl contents of 2.9, 6.1, and 26.0 m...

In order to characterize possible disulfide-linked interactions between lens fiber cell membranes and crystallins, two-dimensional diagonal electrophoresis has been used in combination with Western blot analysis. When these blots were probed with monospecific antisera against alpha, beta and gamma crystallins, membrane from five individual normal lenses showed no disulfide-bonded components. Me...

A cloned cell line was derived from a culture of Nakano mouse lens epithelial cells. The cloned cells grew vigorously and produced large numbers of lentoid bodies. Sodium dodecyl sulfate (SDS) and non-SDS slab-gel electrophoresis of the soluble proteins from the cultured cell revealed protein bands identical in pattern to those of purified gamma crystallin. Antibody to mouse gamma crystallin re...

Water proton relaxation studies were performed on total soluble protein (TSP) solutions and individual alpha-, beta-, and gamma-crystallin fractions derived from weanling rat lenses, 16 month, and 60- to 70-year-old normal human lenses. The effects of perturbants (temperature and acrylamide) were monitored in order to delineate further the mechanisms involved in the generation of the cold catar...

PURPOSE The photobiology of purified recombinant crystallins has not been studied. Here we examine photo-induced aggregation of purified recombinant mouse betaA3-crystallin (rbetaA3) and compare it with that of betaL-crystallins isolated from bovine lenses. METHODS rbetaA3-Crystallin was expressed in baculovirus-infected Sf9 cells and purified by ion-exchange and gel-filtration chromatography...

PURPOSE Crystallins are expressed at high levels in lens fiber cells. Recent studies have revealed that several members of the alpha, beta, and gamma-crystallin family are also distributed in many non-lens tissues, though at lower levels. We observed that the use of retinal RNA as target for both custom I-Gene microarrays and Affymetrix GeneChips revealed significant expression of many crystall...

PURPOSE To study the comparative structural and functional changes between wild-type (wt) and N-terminal congenital cataract causing αA-crystallin mutants (R12C, R21L, R49C, and R54C) upon exposure to different dosages of gamma rays. METHODS Alpha A crystallin N-terminal mutants were created with the site-directed mutagenesis method. The recombinantly overexpressed and purified wt and mutant ...