Transduced protein transduction domain linked HSP27 protected LECs against UVB radiation-induced damage.
PTD-fusion protein technology was used to transduce heat shock protein 27 (HSP27), an anti-apoptotic protein, into human lens epithelial cells (HLECs) (SRA01/04). The protein transduction domain (PTD) of the 11-amino acid YGRKKRRQRRR was tagged at the N-terminus of HSP27. The fusion protein was purified from bacteria transformed with a pKYB-PTD-HSP27 construct. The HLECs were incubated with PTD-HSP27-FITC and the fluorescence inside HLECs was found by fluorescence microscopic examination. To test the ability of PTD-HSP27 to pass through the corneas, PTD-HSP27-FITC was dropped onto the conjunctival sacs of rabbits; fluorescent labeled PTD-HSP27 was then observed in the rabbit aqueous humor. After being incubated with the PTD-HSP27 protein and irradiated with ultraviolet-B (UVB) light, HLECs was analyzed by flow cytometry, Hoechst 33258 staining and measurement of the potential of the mitochondrial transmembrane. HLECs incubated with PTD-HSP27 had a lower apoptotic rate and a higher mitochondrial membrane potential than the control cells. PTD-HSP27 appears to be sufficient to protect HLECs against UVB-induced apoptosis.
Department of Ophthalmology, Affiliated First Hospital of Jinan University, 613 Huangpu Road, Guangzhou, Guangdong, China.
Lian
L
Liu
Biomedical Institute of Jinan University, Jinan University, Guangzhou, Guangdong, China.
Rongjie
R
Yu
Department of Ophthalmology, Affiliated First Hospital of Jinan University, 613 Huangpu Road, Guangzhou, Guangdong, China.
Yuehuan
Y
Shi
Biomedical Institute of Jinan University, Jinan University, Guangzhou, Guangdong, China.
Yun
Y
Dai
Biomedical Institute of Jinan University, Jinan University, Guangzhou, Guangdong, China.
Zhixing
Z
Zeng
Biomedical Institute of Jinan University, Jinan University, Guangzhou, Guangdong, China.
Xiaoling
X
Guo
Department of Ophthalmology, Affiliated First Hospital of Jinan University, 613 Huangpu Road, Guangzhou, Guangdong, China.
Qingshan
Q
Ji
Department of Ophthalmology, Affiliated First Hospital of Jinan University, 613 Huangpu Road, Guangzhou, Guangdong, China.
Guifang
G
Wang
Department of Ophthalmology, Affiliated First Hospital of Jinan University, 613 Huangpu Road, Guangzhou, Guangdong, China. Electronic address: zjx85221206@126.com.
Jingxiang
J
Zhong
D000818:Animals; D000867:Anterior Chamber / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects / Q000528:radiation effects; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D004847:Epithelial Cells / Q000378:metabolism / Q000528:radiation effects*; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D015870:Gene Expression / Q000502:physiology; D015316:Genetic Therapy; D055551:HSP27 Heat-Shock Proteins / Q000235:genetics*; D006360:Heat-Shock Proteins; D006801:Humans; D007908:Lens, Crystalline / Q000378:metabolism / Q000528:radiation effects*; D008297:Male; D008564:Membrane Potentials; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000378:metabolism; D018832:Molecular Chaperones; D010446:Peptide Fragments / Q000235:genetics*; D017434:Protein Structure, Tertiary; D011817:Rabbits; D011833:Radiation Injuries, Experimental / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014161:Transduction, Genetic; D014466:Ultraviolet Rays; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000235:genetics*
D055551:HSP27 Heat-Shock Proteins; C057725:HSPB1 protein, human; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D054322:tat Gene Products, Human Immunodeficiency Virus; C497137:tat peptide (47-57), Human immunodeficiency virus 1; D016650:Fluorescein-5-isothiocyanate
HSP27; apoptosis; cataract; human lens epithelial cell (HLEC); protein transduction domain (PTD); transactivator of transcription (TAT)