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Protamine interaction with the epithelial cell surface.

M W Peterson1, D Gruenhaupt

  • 1Department of Medicine, College of Medicine, University of Iowa, Iowa City 52242.

Journal of Applied Physiology (Bethesda, Md. : 1985)
|January 1, 1992
PubMed
Summary
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Protamine (PRO) binding to kidney cells is specific and occurs in two phases. Trypsin treatment blocks PRO binding and cell injury, indicating protein interactions are key.

Area of Science:

  • Cell Biology
  • Biochemistry
  • Epithelial Physiology

Background:

  • Polycation protamine (PRO) alters Madin Darby canine kidney (MDCK) cell function, increasing short-circuit current and decreasing barrier integrity.
  • Previous studies indicated PRO affects epithelial cell function, but the interaction mechanism at the cell surface was unclear.

Purpose of the Study:

  • To investigate the specific binding of protamine (PRO) to the surface of Madin Darby canine kidney (MDCK) cells.
  • To elucidate the role of cell surface components in PRO binding and subsequent cellular effects.

Main Methods:

  • Protamine (PRO) was labeled with carbon-14 ([14C]) using reductive alkylation for binding studies.
  • Competition assays were performed using unlabeled PRO, polyanions, serum, albumin, neuraminidase, heparinase, and trypsin.

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  • Cellular effects, including mannitol permeability and transepithelial electrical resistance, were measured.

    • Main Results:

    • [14C]PRO binding to MDCK cells occurred in a biphasic pattern, with 20% binding within 30 minutes.
    • PRO binding demonstrated specificity, inhibited by unlabeled PRO, polyanions, serum, and albumin.
    • Neuraminidase and heparinase treatments did not inhibit PRO binding or protect cells from PRO-induced injury, despite altering cell surface components.
    • Trypsin treatment significantly reduced both PRO binding and the increase in mannitol permeability.

    Conclusions:

    • Protamine (PRO) binds specifically to proteins on the surface of Madin Darby canine kidney (MDCK) cells.
    • Cell surface proteins, rather than sialic acid or heparan sulfate, mediate the interaction between PRO and MDCK cells.
    • The interaction with cell surface proteins is critical for PRO-induced changes in epithelial barrier function.