Susceptibility to oxidative stress: proteomic analysis of bronchoalveolar lavage from ozone-sensitive and ozone-resistant strains of mice.
Previous studies have shown that the pulmonary response to ozone (O(3)) varies greatly among strains of mice, but the factor(s) and the mechanism(s) that are responsible for this differential susceptibility have not yet been clearly identified. The present study explores the molecular bases for this differential O(3) susceptibility by studying the expression of proteins associated to the epithelial lining fluid (ELF) from two strains of mice, C57BL/6J and the C3H/HeJ, respectively described as O(3)-sensitive and O(3)-resistant. The ELF proteins of these two strains were displayed by two-dimensional gel electrophoresis (2-DE) of bronchoalveolar lavage fluids (BALFs) and the protein patterns obtained with BALF samples of both strains were compared. Two major differences were observed between the BALF 2-DE protein maps obtained from C57BL/6J and C3H/HeJ strains. First, two isoforms of the antioxidant protein 2 (AOP2) were detected in a strain-dependent manner: C3J/HeJ possesses only AOP2a (isoelectric point 5.7) and C57BL/6J exhibits only AOP2b (isoelectric point 6.0). Second, the levels of anti-inflammatory and immunosuppressive Clara cell protein-16 (CC16) were 1.3 times higher in the BALF from resistant C3H/HeJ than from sensitive C57BL/6 mice. Moreover, two 6 kDa isoforms of CC16 with isoelectric points of 4.9 (CC16a) and 5.2 (CC16b) are detected in both strains. Interestingly, the C57BL/6J strain had a twice decreased level of the acidic isoform of CC16 compared to C3H/HeJ. Our results suggest that AOP2 and CC16 might participate in the protection of the pulmonary tract to O(3)-induced lung injury. The possible differential contribution of specific protein isoforms in the differential susceptibility to oxidative stress is discussed.
Department of Biological Chemistry, University of Mons-Hainaut, Mons, Belgium. ruddy.wattiez@umh.ac.be
Ruddy
R
Wattiez
Isabelle
I
Noël-Georis
Christine
C
Cruyt
Fabrice
F
Broeckaert
Alfred
A
Bernard
Paul
P
Falmagne
D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D000975:Antioxidants / Q000302:isolation & purification; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry*; D005260:Female; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D018384:Oxidative Stress; D010126:Ozone / Q000633:toxicity*; D010544:Peroxidases; D054465:Peroxiredoxin VI; D054464:Peroxiredoxins; D011506:Proteins / Q000235:genetics / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods*; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
D000975:Antioxidants; D011506:Proteins; D010126:Ozone; D010544:Peroxidases; D054465:Peroxiredoxin VI; D054464:Peroxiredoxins; C515983:Prdx6 protein, mouse