The presence and properties of myocilin in the aqueous humor.
OBJECTIVE
To determine whether myocilin is present in the aqueous humor (AH) and to examine certain properties of this protein.


METHODS
Human AH was obtained at the time of either glaucoma surgery or cataract extraction. Monkey AH was obtained at the time of death, and bovine aqueous was obtained from eyes delivered from an abattoir. Column chromatography was performed on aqueous samples to determine the approximate size of the myocilin present. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analysis were performed using antibody prepared against a peptide sequence in myocilin. Analysis of the bovine proteins present in AH that were retained by a microporous filter was also performed using western blot analysis.


RESULTS
By western blot analysis, myocilin was present in human, monkey, and bovine AH. The apparent molecular size of the myocilin present in the AH were greater than 250,000 Da, when quantified with a gel filtration column. Myocilin appeared to be hydrophobic and was one of the proteins that was retained on microporous filters that were obstructed by AH.


CONCLUSIONS
Myocilin is a constituent in the AH. It appears that myocilin is a hydrophobic protein that may exist in an oligomeric state or in association with other proteins. Myocilin is retained by microporous filters and may be involved in the obstruction of these filters that occurs when AH is perfused through them.
National Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA. russellp@intra.nei.nih.gov
P
P
Russell
E R
ER
Tamm
F J
FJ
Grehn
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G
Picht
M
M
Johnson
D000818:Animals; D001082:Aqueous Humor / Q000378:metabolism*; D015153:Blotting, Western; D002386:Cataract / Q000150:complications / Q000378:metabolism; D002387:Cataract Extraction; D002417:Cattle; D002850:Chromatography, Gel; D003598:Cytoskeletal Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D005136:Eye Proteins / Q000378:metabolism*; D005901:Glaucoma / Q000150:complications / Q000378:metabolism / Q000601:surgery; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D008251:Macaca; D008970:Molecular Weight; D000099179:Myocilin
D003598:Cytoskeletal Proteins; D005136:Eye Proteins; D006023:Glycoproteins; D000099179:Myocilin