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Monica Berry, Roger B. Ellingham, Anthony P. Corfield; Membrane-Associated Mucins in Normal Human Conjunctiva. Invest. Ophthalmol. Vis. Sci. 2000;41(2):398-403.
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purpose. To examine the presence of specific membrane-associated mucins in
normal human conjunctiva.
methods. Glycoconjugates were extracted from membranes with two detergents:
octylglucoside and Triton X114. Mucins were separated by cesium
chloride density gradient centrifugation. Size was assessed by gel
filtration on Sepharose CL2B and charge by ion-exchange chromatography
on MonoQ. Cross reaction with antibodies against mucin gene products
was assessed in blots of electrophoresis gels.
results. Extraction of total tissue membranes yielded material with a buoyant
density typical of mucins. Gel filtration showed material reacting with
antimucin antibodies in a range of molecular sizes. Agarose
electrophoresis confirmed the presence of MUC1 and MUC4 and the absence
of MUC2 or MUC5AC. Isolation of membrane mucins by sequential,
exhaustive extraction with octylglucoside followed by Triton X114
suggested the existence of mucins in different membrane environments.
Reagents to carbohydrate epitopes revealed high mobility material,
comigrating with MUC1 and MUC4. Low mobility membrane-bound mucins did
not cross-react with any antibodies to mucin genes known to be
expressed in human conjunctiva.
conclusions. Membrane-associated mucins are distinct from secreted mucins in normal
human conjunctiva. MUC1 and MUC4 mature products decorate the membranes
of conjunctival epithelial cells. Their segregation between octyl
glucoside and the detergent and aqueous phases of Triton X114 suggests
a variety of membrane anchoring modes.
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