Analysis of human ocular mucus: effects of neuraminidase and chitinase enzymes

Cornea
Pablo ArgüesoT Girbés

Abstract

Our goal was to establish the characteristic migration pattern on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of high molecular weight mucins from human ocular mucus and the effects of treatment with exo- and endoglycosidases. Chromatography by gel filtration with Sepharose CL-4B was performed on samples collected from normal subjects. Human ocular mucins from the high molecular weight fraction were digested with exoglycosidases (neuraminidase, N-acetyl-beta-D-glucosaminidase, beta-D-glucosidase) and endoglycosidases (chitinase, lysozyme); and the resulting products were analyzed by electrophoresis. Carbohydrate identification was performed using lectin probes. The migration of the ocular mucins on SDS-PAGE stopped after treatment with neuraminidase, which removes the terminal negatively charged sialic acid residues from mucin. Chitinase (beta(1-4)N-acetylglucosaminidase) treatment increased the electrophoretic migration of mucins. Staining with wheat germ agglutinin and Maackia amurensis agglutinin lectins showed that these mucins contain beta(1-4)NAcGlc and SAa(2-3)Gal linkages. These studies demonstrate that the mobility of human ocular mucins on SDS-PAGE is determined by their intrinsic total negati...Continue Reading

Citations

May 10, 2008·Glycoconjugate Journal·Louise RoyleS D Carrington
Oct 6, 2001·Survey of Ophthalmology·Margarita Calonge
May 5, 2012·Journal of Ocular Pharmacology and Therapeutics : the Official Journal of the Association for Ocular Pharmacology and Therapeutics·William G GensheimerJames V Aquavella
Sep 28, 2002·Cornea·Seung Eun JungEung Kweon Kim
Jan 25, 2008·Cornea·Maria MusumeciSalvatore Musumeci
Jun 11, 2009·Cornea·Maria MusumeciSalvatore Musumeci
Oct 4, 2013·Expert Opinion on Investigational Drugs·Murat DogruKazuo Tsubota
Jul 23, 1999·Biotechnic & Histochemistry : Official Publication of the Biological Stain Commission·M Kurpakus WheaterL D Hazlett
Jun 27, 2006·Progress in Histochemistry and Cytochemistry·F Paulsen, M Berry
Dec 9, 2004·Chemical Reviews·M N V Ravi KumarA J Domb
Aug 25, 2001·Experimental Eye Research·Pablo Argüeso, Ilene K Gipson
Oct 10, 2012·Carbohydrate Polymers·Natália R PiresJudith P A Feitosa

Related Concepts

Sepharose CL 4B
Teens
Endochitinase
Molecular Sieve Chromatography
SDS-PAGE
Lacrimal Gland Structure
Mucins
Mucous Body Substance
Neuraminidase
Polysaccharides

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