Chemical characterisation of bovine conjunctival, mucin spfcies

C. Morris*, M. S. Ball, N. H. Facker

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

Purpose; To identify the species of mucin produced by bovine conjunctiva and to characterise their protein and sugar content by chemical analysis. Methods: The mucins were purified from a bovine conjunctival extract by a single step size exclusion clironialography purification using Superose 12 {Pharmacia) and high salt/reducing conditions. Individual species were separated by polyacryl amide/agarose composite gel eleclrophoresis and/or polyethylene glycol modified SDS-PAuE. Amino acid analysis was performed using gaseous HC1 hydrolysis followed by f-MOC derivatisation (Aminomale, GBC). Monosaccharides and oligosaccharides were separated and analysed by high performance anion exchange chromatography (Dionex). Peptides were produced by digestion with the serine protease R141A (Thermits sp.) and separated by size exclusion and reverse phase HPLC. A GlycoSite (Beckman) automated sequenator was used to perform the Edman sequencing of peptides. Results: Several species of mucin (at least three) were recovered from the conjunctival surface and arnino acid analysis showed that they are lower in total serine and threonine than mucins described from other sources. Also, there is more serine than threonine in these proteins, wliich is in concert with oilier published results on ocular mucins1. The monosaccharide pattern is similar to that found in other mucins, but is also rich in mannose, suggesting uie possibility of a greater N-linked oligosaccharides. Conclusion: Mucin species from bovine conjunctiva have been purified and separated and compositional analysis of amino acids and carbohydrates completed. Further analysis by Edman sequencing of the glycopeptides. and oligosaccharide fine structure analysis may enable the mucin class identification, taking into account species differences.

Original languageEnglish
JournalInvestigative Ophthalmology and Visual Science
Volume38
Issue number4
Publication statusPublished - 1997

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