Expression of a copper-containing amine oxidase by human ciliary body.
OBJECTIVE
To examine the molecular structure and ultrastructural distribution of a novel amine oxidase in human ciliary body.


METHODS
Human ciliary bodies were solubilized with a nonionic detergent. The solubilized material was subjected to affinity chromatography with 2B4.14.1, a monoclonal antibody which recognizes a family of ciliary body glycoproteins. Proteins eluted from the affinity column were further separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Peptides produced from a 2B4.14. 1-reactive protein with an approximate molecular weight of 100 kDa were analyzed by Edman degradation. The protein thus identified was further examined by Western blotting and immunoelectron microscopy with anti-peptide antisera.


RESULTS
Peptide sequences from the 100 kDa ciliary body protein were identical to the predicted protein sequence of an amine oxidase identified recently in a human placental cDNA library. The identity of the ciliary body protein was confirmed by Western blotting with rabbit antiserum generated against the predicted carboxy-terminal peptide of human placenta amine oxidase. Western blotting under nonreducing conditions and following glycosidase digestion indicated that the native enzyme is a disulfide-linked homodimer with multiple N-linked oligosaccharide side chains. By immunoelectron microscopy, the ciliary body amine oxidase was localized to the plasma membranes of inner epithelial cells.


CONCLUSIONS
Human placenta amine oxidase is present on the plasma membranes of ciliary body inner epithelial cells. This finding provides a potential explanation for amine oxidase enzyme activity detected in previous studies of anterior segment tissues. Though the functional role of human placenta amine oxidase in the eye is unclear, it may contribute to the production of H2O2 in aqueous humor.
Howell
D N
DN
Department of Pathology, Veterans Affairs Medical Center, Durham, NC 27705, USA. dhowell@acpub.duke.edu
Valnickova
Z
Z
Oury
T D
TD
Miller
S E
SE
Sanfilippo
F P
FP
Enghild
J J
JJ
D000368:Aged; D000369:Aged, 80 and over; D006631:Amine Oxidase (Copper-Containing) / Q000737:chemistry* / Q000378:metabolism*; D000595:Amino Acid Sequence; D015153:Blotting, Western; D002924:Ciliary Body / Q000737:chemistry / Q000201:enzymology*; D003300:Copper; D004591:Electrophoresis, Polyacrylamide Gel; D004848:Epithelium / Q000201:enzymology; D006801:Humans; D008667:Metalloproteins / Q000737:chemistry / Q000378:metabolism; D016253:Microscopy, Immunoelectron; D008875:Middle Aged; D008995:Monoamine Oxidase / Q000737:chemistry* / Q000378:metabolism*
D008667:Metalloproteins; D003300:Copper; D006631:Amine Oxidase (Copper-Containing); D008995:Monoamine Oxidase