Redox proteomic evaluation of oxidative modification and recovery in a 3D reconstituted human skin tissue model exposed to UVB.
OBJECTIVE
Exposure to UV in humans resulting in sunburn triggers a complex series of events that are a mix of immediate and delayed damage mediation and healing. While studies on the effects of UV exposure on DNA damage and repair have been reported, changes in the oxidative modification of skin proteins are poorly understood at the molecular level, despite the important role played by structural proteins in skin tissue, and the effect of the integrity of these proteins on skin appearance and health. Proteomic molecular mapping of oxidation was here applied to try to enhance understanding of skin damage and recovery from oxidative damage and UVB exposure.


METHODS
A redox proteomic-based approach was applied to evaluating skin protein modification when exposed to varying doses of UVB after initial oxidative stress, via tracking changes in protein oxidation during the healing process in vitro using a full-thickness reconstituted human skin tissue model. Bioassays and structural evaluation confirmed that our cultured skin tissues underwent a normal physiological response to UVB exposure.


RESULTS
A set of potential skin marker peptides was generated, for use in tracking skin protein oxidative modification. Exposure to UVB after thermal oxidative stress was found to result in higher levels of skin protein oxidation than a non-irradiated control for up to seven days after exposure. Recovery of the skin proteins from oxidative stress, as assessed by the overall protein oxidation levels, was found to be impaired by UVB exposure. Oxidative modification was largely observed in skin structural proteins.


CONCLUSIONS
Exposure of skin proteins to UVB exacerbates oxidative damage to structural skin proteins, with higher exposure levels leading to increasingly impaired recovery from this damage. This has potential implications for the functional performance of the proteins and inter-related skin health and cosmetic appearance.
Food & Bio-Based Products, AgResearch Ltd, Private Bag 4749, Christchurch 8140, New Zealand.|Riddet Institute, Massey University, Private Bag 11222 Palmerston North 4442, New Zealand.|Biomolecular Interaction Centre, University of Canterbury, Private Bag 4800 Christchurch 8140, New Zealand.|Wine, Food & Molecular Biosciences, PO Box 85084, Lincoln University, Lincoln 7647, New Zealand.
J M
JM
Dyer
Food & Bio-Based Products, AgResearch Ltd, Private Bag 4749, Christchurch 8140, New Zealand.
S R
SR
Haines
Food & Bio-Based Products, AgResearch Ltd, Private Bag 4749, Christchurch 8140, New Zealand.
A
A
Thomas
Food & Bio-Based Products, AgResearch Ltd, Private Bag 4749, Christchurch 8140, New Zealand.
W
W
Wang
Food & Bio-Based Products, AgResearch Ltd, Private Bag 4749, Christchurch 8140, New Zealand.
R J
RJ
Walls
Food & Bio-Based Products, AgResearch Ltd, Private Bag 4749, Christchurch 8140, New Zealand.|Biomolecular Interaction Centre, University of Canterbury, Private Bag 4800 Christchurch 8140, New Zealand.
S
S
Clerens
Food & Bio-Based Products, AgResearch Ltd, Private Bag 4749, Christchurch 8140, New Zealand.
D P
DP
Harland
D006801:Humans; D008954:Models, Biological; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D040901:Proteomics; D012867:Skin / Q000378:metabolism / Q000528:radiation effects*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014466:Ultraviolet Rays
chemical analysis; collagen; oxidation; proteomics; skin physiology/structure