Myocilin, a component of a membrane-associated protein complex driven by a homologous Q-SNARE domain

W. Michael Dismuke, Brian S Mckay, W. Daniel Stamer

Research output: Contribution to journalArticle

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Abstract

Myocilin is a widely expressed protein with no known function; however, mutations in myocilin appear to manifest uniquely as ocular hypertension and the blinding disease of glaucoma. Using the protein homology/analogy recognition engine (Phyre), we find that the olfactomedin domain of myocilin is similar in sequence motif and structure to a six-blade, kelch repeat motif based on the known crystal structures of such proteins. Additionally, using sequence analysis, we identify a coiled-coil segment of myocilin with homology to human Q-SNARE proteins (inset). Using COS-7 cells expressing full-length human myocilin and a version lacking the C-terminal olfactomedin domain, we identified a membrane-associated protein complex containing myocilin by hydrodynamic analysis. The myocilin construct that included the coiled-coil but lacked the olfactomedin domain formed complexes similar to the full-length protein, indicating that the coiled-coil domain of myocilin is sufficient for myocilin binding to the large detergent-resistant complex. In human retina and retinal pigment epithelium, which express myocilin, we detected the protein in a large, sodium dodecyl sulfate-resistant, membrane-associated complex. We characterized myocilin in human tissues as either a 15 S complex with an Mr of 405000-440000 yielding a slightly elongated globular shape similar to that of known SNARE complexes or a 6.4 S dimer with an Mr of 108000. By identifying the Q-SNARE homology within the second coil of myocilin and documenting its participation in a SNARE-like complex, we provide evidence of a SNARE domain-containing protein associated with a human disease.

Original languageEnglish (US)
Pages (from-to)3606-3613
Number of pages8
JournalBiochemistry
Volume51
Issue number17
DOIs
StatePublished - May 1 2012

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Q-SNARE Proteins
Membrane Proteins
SNARE Proteins
Proteins
trabecular meshwork-induced glucocorticoid response protein
Ocular Hypertension
Retinal Pigments
Retinal Pigment Epithelium

ASJC Scopus subject areas

  • Biochemistry

Cite this

Myocilin, a component of a membrane-associated protein complex driven by a homologous Q-SNARE domain. / Dismuke, W. Michael; Mckay, Brian S; Stamer, W. Daniel.

In: Biochemistry, Vol. 51, No. 17, 01.05.2012, p. 3606-3613.

Research output: Contribution to journalArticle

Dismuke, W. Michael ; Mckay, Brian S ; Stamer, W. Daniel. / Myocilin, a component of a membrane-associated protein complex driven by a homologous Q-SNARE domain. In: Biochemistry. 2012 ; Vol. 51, No. 17. pp. 3606-3613.
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