Does a Conformational Change of

Integral Membrane Proteins Influence

the Shape of Red Blood Cells?

I. Bernhardt

University of the Saarland,

Laboratory of Biophysics,
Saarbrücken, Germany
 Aim of the Work

Does a conformational change of the major red cell

membrane protein (band 3) influencing the ratio of the
area of the outer to the inner leaflet of the membrane
affect the resting shape as speculated by Gimsa and
Ried (1995)?

Is it possible to correlate the cross sectional area of

the membrane proteins of the outer membrane surface
with the red cell shape?
Membrane curvature depending on the
protein conformation

Band 3 outward
conformation

Band 3 inward
conformation
 Background (I)
Based on the bilayer-couple hypothesis (Sheetz and
Singer, 1974), the red cell shape is determined by the
area of the outer leaflet in relation to the inner leaflet of
the membrane.

Insertion of amphiphilic compounds into the membrane

or a transversal redistribution of membrane phospho-
lipids leads to a change of the resting shape!

Other known parameters influencing the shape are, e.g.,

the spectrin-membrane interaction and the pH of the
extracellular solution. The addition of substances, which
affect the band 3 mediated transport induce fast shape
changes.
Experimental conditions leading to different shapes

- Physiological (high) ionic strength (HIS) solution, at different pH

- Low ionic strength (LIS) solution

- Band 3 mediated transport inhibitors

- without changing the Band 3 conformation

niflumic acid (NA)

- changing the Band 3 conformation

DIDS, furosemide (FS)
 AFM image of the surface of a human RBC
Discocyte

whole cell plot, 8 x 8 µm high resolution plot, 250 x 250 nm

 AFM image of the surface of a human RBC
Stomatocyte

whole cell plot, 6 x 6 µm high resolution plot, 250 x 250 nm

 AFM image of the surface of a human RBC
Echinocyte

whole cell plot, 5 x 5 µm high resolution plot, 250 x 250 nm

Mean values for all obtained volumes of the external
part of membrane proteins under conditions leading
to different shapes
30.00

25.00

20.00
Volume, nm³

15.00

10.00

5.00

0.00
Stomatocyte Discocyte Echinocyte

Error bars represent S.E.M., n > 100 for each shape, p < 0.01
 Summary

A change of the volume of the extracellular segment of

membrane proteins in the human erythrocyte (mainly
band 3) and a corresponding change of the cross
sectional area of the protein in the outer half of the
membrane results in an alteration of the ratio of the
external to the internal leaflet area of the membrane
that in turn modifies the shape of the cell.