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 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.
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 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.
Authors: José-Daniel Aroca-Aguilar; Francisco Sánchez-Sánchez; Sikha Ghosh; Ana Fernández-Navarro; Miguel Coca-Prados; Julio Escribano Journal: Invest Ophthalmol Vis Sci Date: 2011-01-05 Impact factor: 4.799
Authors: Joshua T Morgan; Heung Sun Kwon; Joshua A Wood; Dori L Borjesson; Stanislav I Tomarev; Christopher J Murphy; Paul Russell Journal: Exp Eye Res Date: 2015-02-24 Impact factor: 3.467
Authors: Kyle G Howell; Anne M Vrabel; Uttio Roy Chowdhury; William Daniel Stamer; Michael P Fautsch Journal: J Glaucoma Date: 2010-12 Impact factor: 2.503
Authors: Mohamed-Karim Ezzat; Kyle G Howell; Cindy K Bahler; Thomas G Beito; Nils Loewen; Eric M Poeschla; Michael P Fautsch Journal: Exp Eye Res Date: 2008-07-12 Impact factor: 3.467
Authors: Emely A Hoffman; Kristin M Perkumas; Lindsey M Highstrom; W Daniel Stamer Journal: Invest Ophthalmol Vis Sci Date: 2008-10-24 Impact factor: 4.799