Analysis of Acanthamoeba polyphaga surface carbohydrate exposure by FITC-lectin binding and flourescence evaluation

Elloway, E.A.G., Armstrong, Richard A., Bird, R.A., Kelly, S.L. and Smith, S.N. (2004). Analysis of Acanthamoeba polyphaga surface carbohydrate exposure by FITC-lectin binding and flourescence evaluation. Journal of Applied Microbiology, 97 (6), pp. 1319-1325.

Abstract

Aims: Characterization of the representative protozoan Acanthamoeba polyphaga surface carbohydrate exposure by a novel combination of flow cytometry and ligand-receptor analysis. Methods and Results: Trophozoite and cyst morphological forms were exposed to a panel of FITC-lectins. Population fluorescence associated with FITC-lectin binding to acanthamoebal surface moieties was ascertained by flow cytometry. Increasing concentrations of representative FITC-lectins, saturation binding and determination of K d and relative Bmax values were employed to characterize carbohydrate residue exposure. FITC-lectins specific for N-acetylglucosamine, N-acetylgalactosamine and mannose/glucose were readily bound by trophozoite and cyst surfaces. Minor incremental increases in FITC-lectin concentration resulted in significant differences in surface fluorescence intensity and supported the calculation of ligand-binding determinants, Kd and relative B max, which gave a trophozoite and cyst rank order of lectin affinity and surface receptor presence. Conclusions: Trophozoites and cysts expose similar surface carbohydrate residues, foremost amongst which is N-acetylglucosamine, in varying orientation and availability. Significance and Impact of the Study: The outlined versatile combination of flow cytometry and ligand-receptor analysis allowed the characterization of surface carbohydrate exposure by protozoan morphological forms and in turn will support a valid comparison of carbohydrate exposure by other single-cell protozoa and eucaryotic microbes analysed in the same manner.

Publication DOI: https://doi.org/10.1111/j.1365-2672.2004.02430.x
Divisions: Life & Health Sciences > Optometry
Life & Health Sciences > Ophthalmic Research Group
Life & Health Sciences
Life & Health Sciences > Biosciences
Uncontrolled Keywords: acanthamoeba,FITC-lectins,flow cytometry,ligand-receptor analysis,surface carbohydrates,Agricultural and Biological Sciences(all),Applied Microbiology and Biotechnology,Biotechnology,Microbiology
Full Text Link: http://www.wiley.com/bw/journal.asp?ref=1364-5072&site=1
Related URLs: http://www.scopus.com/inward/record.url?scp=9644255754&partnerID=8YFLogxK (Scopus URL)
Published Date: 2004-12-13
Authors: Elloway, E.A.G.
Armstrong, Richard A. ( 0000-0002-5046-3199)
Bird, R.A.
Kelly, S.L.
Smith, S.N.

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