Characterization of soluble protein BCP 11/24 from bovine corneal epithelium, different from the principal soluble protein BCP 54.
The water-soluble fraction of bovine corneal epithelium was analysed by polyacrylamide gel electrophoresis in the presence of SDS (SDS-PAGE). Next to the principal soluble protein BCP 54, which has recently been identified as a corneal aldehyde dehydrogenase (ALDH), another abundant protein was observed, which we have denoted BCP 11/24, due to its estimated molecular weight of 11 kDa in SDS-PAGE and 24 kDa in high performance gel filtration under non-denaturing conditions. This protein was isolated and characterized by biochemical and immunochemical techniques. The isolation of BCP 11/24 was initially hampered by its tendency to bind non-covalently to BCP 54. BCP 11/24 behaves identically in reduced and unreduced SDS-PAGE and is probably not a glycoprotein. Isoelectric focusing indicated microheterogeneity of BCP 11/24, yielding bands with isoelectric points of 6.1, 5.9, 5.7 and 5.6. A rabbit antiserum directed against BCP 11/24, that did not recognize BCP 54, demonstrated that the distribution of BCP 11/24 in different ocular tissues as well as its light microscopic localization in corneal epithelium is strikingly similar to that of BCP 54. Together with its tendency to interact with BCP 54 in vitro, this suggests the possibility that BCP 11/24 is associated with BCP 54 in vivo, fulfilling a function which may be related to the activity of BCP 54 as a corneal ALDH. In contrast with BCP 54, however, BCP 11/24 was not detectable in corneal endothelium. The antiserum did not detect any immunologically related molecules in corneal epithelium extracts of sheep, human or rat origin, indicating that BCP 11/24 is probably not as highly conserved as BCP 54.
Bakker
C
C
Department of Ophthalmo-Immunology, The Netherlands Ophthalmic Research Institute, Amsterdam.
Pasmans
S
S
Verhagen
C
C
Van Haren
M
M
Van der Gaag
R
R
Hoekzema
R
R
D000444:Aldehyde Dehydrogenase; D000818:Animals; D002417:Cattle; D002850:Chromatography, Gel; D003315:Cornea / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D004848:Epithelium / Q000737:chemistry; D005136:Eye Proteins / Q000032:analysis*; D007525:Isoelectric Focusing; D008970:Molecular Weight
D005136:Eye Proteins; C074043:corneal protein 11-24kDa, bovine; C030863:corneal protein 54, bovine; D000444:Aldehyde Dehydrogenase