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The interaction between wheat germ agglutinin and membrane incorporated glycophorin A. An optical binding study

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Abstract

A novel integrated optical technique is used to monitor the kinetics of incorporation of glycophorin A (GPA) from solution into a planar dimyristoylphosphatidylcholine-cholesterol bilayer membrane, and the subsequent binding of wheat germ agglutinin (WGA) to the membrane-incorporated GPA. The technique significantly improves the attainable accuracy of kinetic measurements. The number of bound molecules can be determined to a precision of ca ± 80 mol µm−2. Our results show that GPA incorporates spontaneously into the bilayer. Binding of WGA to GPA is optimal in the presence of human serum albumin, and can be reversed byN-acetyl-d-glucosamine. The kinetics of the binding are consistent with the presence of two classes of kinetically distinguishable binding sites with association rates of 2.0×104 and 9.6×102 M−1 s−1, and dissociation rates of 2.7×10−3 s−1 and <10−5 s−1, respectively. A stoichiometry of 4 WGA monomers per GPA monomer was determined as characteristic of the overall binding interaction.

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Abbreviations

DMPC:

dimyristoylphosphatidylcholine

GlcNAc:

N-acetyl-d-glucosamine

GPA:

glycophorin A

HSA:

human serum albumin

NeuNAc:

N-acetyl-d-neuraminic acid

TE:

transverse electric

TM:

transverse magnetic

WGA:

wheat germ agglutinin

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Authors and Affiliations

  1. Department of Biophysical Chemistry, Biozentrum, CH-4056, Basle, Switzerland

    Jeremy J. Ramsden

  2. Departments of Biochemistry and Molecular Biophysics and Medicinal Chemistry, Medical College of Virginia, Virginia Commonwealth University, 23298-0540, Richmond, VA, USA

    Christine Schubert Wright

Authors
  1. Jeremy J. Ramsden
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  2. Christine Schubert Wright
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Ramsden, J.J., Wright, C.S. The interaction between wheat germ agglutinin and membrane incorporated glycophorin A. An optical binding study. Glycoconjugate J 12, 113–121 (1995). https://doi.org/10.1007/BF00731354

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  • DOI: https://doi.org/10.1007/BF00731354

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