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Abstract #5588 Published in IGR 2-1

Micropurification of beta- and gamma-crystallins from rabbit aqueous humor

Leone MG; Saso L; Cheng CY; Silvestrini B
International Journal of Biological Macromolecules 1999; 26:167-171


Soluble crystallins are normally present in the aqueous humor, originating from the lens, and their concentration may increase in certain conditions such as cataract, possibly contributing to aqueous outflow pathway obstruction, leading to glaucoma. Whether the stability and tendency of aqueous crystallins to aggregate are different in patients with certain forms of open-angle glaucoma has not yet been established, mainly due to the lack of a suitable purification procedure from this fluid in which crystallins are present at very low concentrations together with dozens of other proteins. About 4 μg each of β- and γ-crystallins were obtained from 20 ml of rabbit aqueous humor by C8 reversed-phase high-performance liquid chromatography (HPLC) and high-performance electrophoresis chromatography (HPEC). The identity of the proteins was confirmed by amino acid analysis following sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and electrophoretic blotting onto polyvinylidene fluoride membranes, with or without previous digestion with Staphylococcus aureus protease V8.

Dr. M.G. Leone, Department of Pharmacology of Natural Substances and General Physiology, University of Rome La Sapienza, Rome, Italy


Classification:

2.6 Aqueous humor dynamics (Part of: 2 Anatomical structures in glaucoma)
9.4.4 Glaucomas associated with disorders of the lens (Part of: 9 Clinical forms of glaucomas > 9.4 Glaucomas associated with other ocular and systemic disorders)



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