THE ERYTHROCYTE MEMBRANE

The erythrocyte membrane, because of its easy accessibility, is one of

the best studied biologic membranes. Composed of a lipid bilayer and
an underlying cortical membrane skeleton (Fig. 164-1), the membrane
provides the erythrocyte the deformability and stability required to
withstand its travels through the circulation. In one circulatory cycle
throughout the body, the erythrocyte is subjected to high sheer stress
in the arterial system, dramatic size and shape changes in the
microcirculation with capillary diameters as small as 7.5 m, and
marked fluctuations in tonicity, pH, and PO2.

Figure 164-1 The erythrocyte membrane. A model of the major proteins of the erythrocyte
membrane

Membrane Lipids
Red cell membrane lipids are asymmetrically distributed across the
bilayer membrane, reflecting a steady state involving a constant
exchange of phospholipids between the two bilayer hemileaflets.

Glycolipids and cholesterol are intercalated between the

phospholipids in the bilayer with their long axes perpendicular to the
bilayer plane. Glycolipids, located in the external half of the bilayer
with their carbohydrate moieties extending into the aqueous phase,
carry several important red cell antigens and serve other important
functions. Phospholipids are asymmetrically organized, with the
choline phospholipids, phosphatidylcholine and sphingomyelin,
primarily in the outer half of the bilayer, and the amino
phospholipids, phosphatidylethanolamine and phosphatidylserine
(PS), in the inner half of the bilayer. In pathologic states, such as
thalassemia, sickle cell disease, and diabetes, loss of phospholipid
asymmetry with externalization of PS leads to activation of blood
clotting through conversion of prothrombin to thrombin and
facilitates macrophage attachment to erythrocytes, marking them for
destruction. Mature erythrocytes are unable to synthesize fatty acids,
phospholipids, or cholesterol de novo and depend on lipid exchange
and fatty acid acylation as mechanisms for phospholipid repair and
renewal.

Membrane Proteins
The red cell membrane contains about a dozen major proteins and
hundreds of minor ones. Membrane proteins are classified as integral,
penetrating or crossing the lipid bilayer and interacting with the
hydrophobic lipid core, or peripheral, interacting with integral proteins
or lipids at the membrane surface but not penetrating into the bilayer
core. Integral membrane proteins include the glycophorins, the Rh
proteins, Kell and Duffy antigens, and transport proteins such as band
3 (AE1, anion exchanger 1, SLC4A1), Na+, K+-ATPase, Ca2+-ATPase, and
Mg2+-ATPase. Numerous membrane receptors and antigens are
present on integral membrane proteins. Peripheral membrane
proteins are on the cytoplasmic membrane face and include enzymes
such as glyceraldehyde 3-phosphate dehydrogenase and the structural
proteins of the spectrin-actinbased membrane skeleton.

Integral Membrane Proteins

Band 3, the major integral protein of the red cell, has two primary
functions: ion transport and maintenance of protein-protein
interactions. Band 3 mediates chloride-bicarbonate exchange and
provides a binding site for glycolytic enzymes, hemoglobin, and the
skeletal proteins ankyrin, protein 4.1, and protein 4.2. A single Nglycan chain attached to an Asn in the membrane spanning domain of
band 3 is composed of N-acetyl-D-lactosamine units arranged in an
unbranched, linear fashion in fetal erythrocytes (i antigen) and in a
branched fashion in adult cells (I antigen).
The glycophorins are the next most abundant family of integral
membrane proteins. They provide most of the negative surface charge
required by red cells to avoid sticking to each other and to the vascular
wall. They are involved in transmembrane signaling and carry
receptors for Plasmodium falciparum, a number of viruses and bacteria,
and several blood group antigens. Other integral proteins include the
Rh proteins, Kell antigen, the glucose transporter, the urea
transporter, Na+, K+-ATPase, Ca2+-ATPase, Mg2+-ATPase, various
kinases and phosphatases, acetylcholinesterase, decay accelerating
factor, complement proteins, and receptors for transferrin, insulin,
insulin-like growth factors, thyroid hormone, parathyroid hormone, adrenergic agonists, cholinergic agents, diphtheria toxin,
ceruloplasmin, and opiates.

Peripheral Membrane Proteins

Spectrin is the major component of the membrane skeleton. It is
composed of two subunits, and spectrin, that are structurally
related but functionally distinct. These subunits intertwine around
each other in an antiparallel fashion to form heterodimers, which in
turn self-associate head-to-head to form tetramersslender, twisted
wormlike molecules that extend to a total length of about 100 nm.
Spectrin is highly flexible and assumes a variety of conformations, an
unusual property that may be critical for normal membrane pliancy.
The spectrin-based membrane skeleton is linked to the plasma
membrane through the actin-protein 4.1 junctional complex, through
spectrin-ankyrin interactions, and through binding of a multiprotein

complex containing Rh proteins, Rh-associated glycoproteins, CD47,

LW, glycophorin B, and protein 4.2 to ankyrin. Protein 4.1, a protein
necessary for normal membrane stability, interacts with spectrin,
actin, and other proteins of the red cell membrane. Ankyrin serves as
the primary linkage protein for the high-affinity binding of spectrin to
the inner membrane through interactions with the cytoplasmic
domain of band 3. Protein 4.2 is a peripheral membrane protein that
helps link the skeleton to the lipid bilayer through interactions with
ankyrin and band 3. Other integral proteins include actin and the
actin-associated proteins, the adducins, dematin, myosin,
tropomyosin, and tropomodulin.
Erythrocyte membrane disorders result from alterations in the
quantity or quality (or both) of individual proteins and their dynamic
interactions with each other. Disruption of the vertical protein-protein
interactions of the membrane, that is, the spectrin-ankyrinband 3
linkage or the band 3protein 4.2 interaction, leads to uncoupling of
the membrane skeleton from the lipid bilayer. This leads to membrane
instability with loss of lipids and some integral membrane proteins,
resulting in loss of membrane surface area and the phenotype of
spherocytosis. Disruption of the horizontal interactions of membrane
skeleton proteins, including perturbation of spectrin self-association
or junctional complex protein-protein interactions, leads to
membrane instability, altered membrane deformability and
mechanical properties, and the phenotype of elliptocytosis.