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Michael P. Fautsch, Douglas H. Johnson; Characterization of Myocilin–Myocilin Interactions. Invest. Ophthalmol. Vis. Sci. 2001;42(10):2324-2331.
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© ARVO (1962-2015); The Authors (2016-present)
purpose. To determine whether myocilin (MYOC; also referred to as TIGR) is
present as a complex in human aqueous humor, whether part of the
complex formation may be due to MYOC–MYOC interactions and to
characterize the sites of interaction.
methods. Human aqueous humor was analyzed by using a gel filtration column for
the identification of MYOC complexes. MYOC–MYOC interactions were
studied with a yeast two-hybrid system. Expression of full-length and
truncated MYOC proteins in AH109 yeast was analyzed for growth and
color on minimal medium. Site-directed mutagenesis was used to
selectively mutate eight leucine residues within the leucine zipper
motif. In vitro transcription and translation was used to verify yeast
results. MYOC was found to be present in human aqueous humor as a complex
ranging from 120 to 180 kDa. Expression of full-length MYOC in yeast as
well as in vitro binding studies revealed that MYOC can interact with
itself. MYOC–MYOC interactions occurred mainly within amino acids
117-166, a region containing a leucine zipper domain. Glycine
substitution for selective leucine residues confirmed that MYOC–MYOC
interactions occurred mainly within the leucine zipper domain.
conclusions. MYOC is present in human aqueous humor, not as a monomer but as a
complex. Part of this complex may form due to MYOC–MYOC interactions
that take place mainly within the leucine zipper
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