Spectroscopic analysis of lens recombinant betaB2- and gammaC-crystallin.
OBJECTIVE
To compare the spectroscopic and unfolding properties of human lens [beta]B2- and [gamma]C-crystallin with those of [alpha]A-crystallin.


METHODS
Human lens [beta]B2- and [gamma]C-crystallin were cloned and measured spectroscopically. The unfolding curves in response to guanidine HCl (GdnHCl) and heat were also obtained by measuring Trp fluorescence emission intensity or emission maximum wavelength with increasing perturbation.


RESULTS
Very similar spectroscopic and unfolding properties were seen with [beta]B2- and [gamma]C-crystallin, but both demonstrated great differences compared with [alpha]A-crystallin. Unlike [alpha]A-crystallin, [beta]B2- and [gamma]C-crystallin showed very little binding to Bis-ANS (4,4'-dianilino-1,1'-binaphthalene-5,5'-disulfonic acid), a hydrophobic fluorescence probe. Both [beta]B2- and [gamma]C-crystallin were more resistant than [alpha]A-crystallin to GdnHCl-induced unfolding, but [alpha]A-crystallin was more resistant than [beta]B2- and [gamma]C-crystallin to heat induced unfolding.


CONCLUSIONS
It was observed that [beta]B2- and [gamma]C-crystallin showed more similar spectroscopic and unfolding properties with each other than each of them showed with [alpha]A-crystallin.
Fu
L
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Center for Ophthalmic Research, Brigham and Women's Hospital, Department of Ophthalmology, Harvard Medical School, Boston, MA 02115, USA.
Liang
J J
JJ
D002942:Circular Dichroism; D003459:Crystallins / Q000737:chemistry* / Q000235:genetics; D017931:DNA Primers / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006358:Hot Temperature; D006801:Humans; D007908:Lens, Crystalline / Q000737:chemistry; D016133:Polymerase Chain Reaction; D011487:Protein Conformation; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D003459:Crystallins; D017931:DNA Primers; D011994:Recombinant Proteins