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Yingying Yu, Yongzhen Bao; Effect of UV-B Irradiation on the Secondary Structural Properties pior to its Chaperone-like Activity. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):5582.
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To evaluate the damaging effect of UV-B irradiation on the secondary structural properties of α-crystallin and its chaperone-like activity.
α-crystallin was isolated from bovine lenses using fast protein liquid chromatography. The purified α-crystallin was subjected to UV-B irradiation (308 nm; 67mW/cm2; 2, 4, 8, 12, 24, 36, 48, 60h, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13d). We measured the Fourier transform infrared (FTIR) spectroscopy in amide I band (1600－1700 cm-1) and the chaperone activity of both irradiated and non-irradiated α-crystallins.
The FTIR spectrum showed that the secondary structure of α-crystallin altered with increasing irradiation time, and almost all of the intramolecular anti-parallel β-sheet structures were lost at time above 48h，leading to the formation of random coil structures. The chaperone activity of α-crystallin with the irradiation time fewer than 60h remained intact. However, this activity was reduced to 70% after irradiation time at 3d and decreased with increasing doses of UV-B irradiation. Moreover, this activity was reduced to 30% after irradiation time over 6d and remained constant.
Our study suggests that the intramolecular anti-parallel β-sheet structures of α-crystallin are susceptible to the damaging effect of UV-B irradiation, leading to the subsequent decrease of its chaperone-like activity.
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