The presence and properties of myocilin in the aqueous humor

PURPOSE. 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 ca taract extraction. Monkey AH was obtained at the time of death, and bovine aqueous was obtained from eyes delivered from an abattoir. Column chromatog raphy was performed on aqueous samples to determine the approximate size of the myocilin present. Sodium dodecyl sulfate-polyacrylamide gel electropho resis (SDS-PAGE) and western blot analysis were performed using antibody pr epared against a peptide sequence in myocilin. Analysis of the bovine prote ins present in AH that were retained by a microporous filter was also perfo rmed using western blot analysis. RESULTS. By western blot analysis, myocilin was present in human, monkey, a nd 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 ma;v exist in an oligomeric state or in assoc iation with other proteins. Myocilin is retained by microporous filters and may be involved in the obstruction of these filters that occurs when AW is perfused through them.