Temperature induced structural changes of beta-crystallin and sphingomyelin binding.
The study of the binding of alpha-crystallin to membranes is potentially important for understanding the function of alpha-crystallin in the ocular lens and the formation of cataracts. Using fluorescence probes, N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-1,2-dihexadecanoyl-sn-glycero-3 -phosphoethanolamine, triethylammonium salt (NBD-PE) and (1,1'-bi(4-anilino)naphthalene-5,5'-disulfonic acid, dipotassium salt (bis-ANS), the temperature dependence of the binding of alpha-crystallin to sphingomyelin liposomes, and the structural changes of alpha-crystallin and sphingomyelin induced by temperature were studied. The influence of the binding of alpha-crystallin on the mobility of the head group region of liposomes of sphingomyelin was dependent on the thermal history of alpha-crystallin. Binding of alpha-crystallin to sphingomyelin caused a decrease in the anisotropy of the fluorophore NBD-PE at or below 37 degrees C. However, when alpha-crystallin or the mixture of alpha-crystallin/sphingomyelin were preincubated near the secondary structure phase transition temperature of 60 degrees C, an increase of the anisotropy of NBD-PE (decrease of lipid head group mobility) was observed when measured at 22 degrees C or 37 degrees C. An inflection near 47 degrees C in the curve of fluorescence anisotropy of bis-ANS pre-incorporated into the alpha-crystallin corresponded to a 3 degrees or 4 degrees structural change of alpha-crystallin. alpha-Crystallin either increases or decreases the flexibility of the head group of sphingomyelin liposomes depending on its structure.
Department of Ophthalmology and Visual Science, University of Louisville, KY 40202, USA.
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D000818:Animals; D016880:Anisotropy; D002417:Cattle; D003459:Crystallins / Q000737:chemistry* / Q000378:metabolism; D005456:Fluorescent Dyes; D006358:Hot Temperature; D008081:Liposomes; D010714:Phosphatidylethanolamines; D011485:Protein Binding; D013050:Spectrometry, Fluorescence; D013109:Sphingomyelins / Q000378:metabolism*
D003459:Crystallins; D005456:Fluorescent Dyes; D008081:Liposomes; D010714:Phosphatidylethanolamines; D013109:Sphingomyelins; C042077:N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)phosphatidylethanolamine