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      Alpha-Crystallin/Lens Lipid Interactions Using Resonance Energy Transfer

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          Abstract

          Resonance energy transfer was used to study the interaction of α-crystallin with lens cortex lipid vesicles. The binding of α-crystallin to cortex lipid vesicles and the preincubation temperature dependence of the binding were confirmed. In this study, the tryptophan of α-crystallin was used as the energy donor, and the fluorescence probe N-(5-dimethylaminonaphthalene-1-sulfonyl)-1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine triethylammonium salt (dansyl DHPE) was chosen as the energy acceptor. Lens cortex lipid vesicles were preincorporated with dansyl DHPE. Energy transfer from the tryptophan of α-crystallin to dansyl DHPE was found and the energy transfer efficiency was calculated. There was a higher energy transfer efficiency between α-crystallin and dansyl DHPE when α-crystallin was preincubated at 65°C compared to 22°C. Data confirmed the binding of α-crystallin to lens cortex lipid and showed that α-crystallin bound more closely to the surface of cortex vesicles when it was preincubated at a higher temperature. This is probably due to the exposure of hydrophobic surfaces when α-crystallin is preincubated at a higher temperature.

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          Subunit Exchange of αA-Crystallin

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            Structural characterization of lipid membranes from clear and cataractous human lenses.

            Lipid composition related structural changes in human cataractous lenses was explored by characterizing the hydrophobic hydrocarbon chains in lipid membranes corresponding to twelve Indian cataractous lenses and eight American clear lenses of similar age. The nuclear-lipid phase transitions corresponding to the clear lenses exhibited significantly higher average transition temperatures (nucleus 33 degrees C, cortex 26.3 degrees C) and cooperativities, 38.1, as compared to the value of 24.1 for the cortical-lipid phase transitions. At 36 degrees C, the phase transitions corresponding to cortical and nuclear lipids indicate a similar degree of disorder, 63%, in the hydrocarbon chains, i.e., similar relative amounts of gauche and trans rotomers. The twelve cataractous lenses investigated all had nuclear opacities, four were brunescent and four had cortical opacities. No significant differences were observed in the phase transition parameters (temperature, cooperativity, magnitude, enthalpy) evaluated for the nuclear-lipid membranes corresponding to the different types of cataracts. Furthermore, for the cataractous membranes, the phase transition parameters obtained for the nuclear lipids were comparable to those evaluated for the cortical lipid membranes. However, the cortical lipids exhibited the highest order in membranes from nuclear cataracts without cortical opacity. The cortical lipids from clear, non-cataractous lenses had the lowest level of order. At 36 degrees C, the degree of order in the cortical lipid from clear lenses was comparable to that from nuclear cataractous lenses without cortical opacity. The transition temperature, and cooperativity were significantly higher for cortical lipids from cataractous lenses as compared to those from clear lenses. At 36 degrees C, the degree of order in the cortical lipid membranes was lower for all cataract types vs. clear lens fractions. Our results suggest the possibility that lipid-lipid interactions could be different in cataractous lens membranes. Lipid compositional and chemical differences must account for these altered lipid interactions. These studies will provide a basis for studying lipid-protein interactions and structure-function relationships in the lens membrane.
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              Author and article information

              Journal
              ORE
              Ophthalmic Res
              10.1159/issn.0030-3747
              Ophthalmic Research
              S. Karger AG
              0030-3747
              1423-0259
              1999
              December 1999
              30 September 1999
              : 31
              : 6
              : 452-462
              Affiliations
              Departments of aOphthalmology and Visual Science and bChemistry, University of Louisville, Ky., USA
              Article
              55571 Ophthalmic Res 1999;31:452–462
              10.1159/000055571
              10474075
              © 1999 S. Karger AG, Basel

              Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher. Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug. Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

              Page count
              Figures: 7, Tables: 1, References: 35, Pages: 11
              Categories
              Original Paper

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