Transport Through Membrane

Transport are of two types

. Passive Transport . Active Transport

Passive transport involves carriers, channels, or direct diffusion

through a membrane.
This type of transport always operates from regions of greater
concentration to regions of lesser concentration.
No external source of energy is required.

Examples of passive transport include

·Simple diffusion
·Channel diffusion
·Facilitated diffusion

There are two types of active transport:

·Primary active transport
·Secondary active transport.

In active transport it is possible to go against the concentration

gradient.
In active transport, a source of energy is required to move the
carrier and its materials.

Primary Active Transport directly uses ATP. An example of primary

active transport is the sodium pump.

Secondary active transport does not directly use ATP.

It takes advantage of a previously existing concentration gradient
(via carriers).

An example of the body needing to move against the concentration

gradient
can be found in looking at glucose in the intestine.

The body has to move glucose against the concentration gradient.

There are three types of transport proteins: channels, carriers, and

pumps
Glucose Transport Through
Intestinal Membrane
Introduction
Intestinal hexose absorption and gluconeogenesis have been
studied in relation to refeeding after two different fasting
phases: a long period of protein sparing during which energy
expenditure is derived from lipid oxidation (phase II), and a
later phase characterized by a rise in plasma corticosterone
triggering protein catabolism (phase III). Such a switch in body
fuel uses, leading to changes in body reserves and
gluconeogenic precursors, could modulate intestinal
gluconeogenesis and glucose transport. The gene and protein
levels, and the cellular localization of the sodium–glucose
cotransporter SGLT1, and of GLUT5 and GLUT2, as well as that
of the key gluconeogenic enzymes phosphoenolpyruvate
carboxykinase (PEPCK) and glucose-6-phosphatase (Glc6Pase)
were measured. PEPCK and Glc6Pase activities were also
determined. In phase III fasted rats, SGLT1 was up-regulated
and intestinal glucose uptake rates were higher than in phase
II fasted and fed rats. PEPCK and Glc6Pase mRNA, protein
levels and activities also increased in phase III. GLUT5 and
GLUT2 were down-regulated throughout the fast, but
increased after refeeding, with GLUT2 recruited to the apical
membrane. The increase in SGLT1 expression during phase III
may allow glucose absorption at low concentrations as soon
as food is available. Furthermore, an increased epithelial
permeability due to fasting may induce a paracellular
movement of glucose. In the absence of intestinal GLUT2
during fasting, Glc6Pase could be involved in glucose release
 to the bloodstream via membrane trafficking. Finally,
refeeding triggered GLUT2 and GLUT5 synthesis and apical
recruitment of GLUT2, to absorb larger amounts of hexoses.

Glucose transport by intestinal epithelial cells A transporter in the

apical domain of the plasma membrane is responsible for the active
uptake of glucose (by cotransport with Na+) from the intestinal
lumen. As a result, dietary glucose is absorbed and concentrated
within intestinal epithelial cells. Glucose is then transferred from
these cells to the underlying connective tissue and blood supply by
facilitated diffusion, mediated by a transporter in the basolateral
domain of the plasma membrane. The system is driven by the Na +-
K+ pump, which is also found in the basolateral domain.

Types Of Pathways
Pores
Pores are one of two routes by which small hydrophilic molecules
(ions) can cross cell (plasma) membranes. These are nonspecific
“holes” in the lipid bilayer, and include movement through the
hydrophilic centers of protein complexes.

Ion Channels
Channels are the second type of pathway by which small
hydrophilic molecules (ions) cross cell membranes. Channels differ
from pores in several ways.

Channels are specific proteins which span the membrane. They can
be purified and studied in isolation.

They exhibit a high order of ionic selectivity which is based on

factors in addition to size. These factors are not well understood.

There are specific drugs which will block specific ion channels at
extremely low concentration. Some very important drugs and
poisons fall into this category.

Channels, unlike pores, are not open all the time. They are
equipped with “gates” which open or close in response to specific
signals. There are two broad classes of channels: ligand gated
channels and voltage gated channels.

Glucose Transporters
The hexoses glucose, galactose and fructose serve as basic fuel
molecules for eucaryotic cells. These molecules are unable to
diffuse across cellular membranes, and require transporter
proteins for entry into and exit from cells. Two distinct groups
of hexose transporters have been identified and classified based
on their dependence on cellular energy:

Transport hexoses down a concentration gradient (GLUT1,

GLUT2, GLUT3, GLUT4 and GLUT5).

Tranport hexoses against a concentration gradient using energy

provided by an electrochemical gradient of sodium, which is
cotransported (SGLUT1).

Each of the transporters has different affinities for glucose and

the other hexoses, which largely dictates their function. GLUT1,
3 and 4 have a high affinity for glucose (Km = 2-5 mM), which
indicates that they are functioning at maximal rate under
physiologic concentrations of glucose (~5 mM). In contrast,
GLUT 2 has a low affinity for glucose (Km ~15 mM), which
allows it to change transport rate in proportion to the
increasing glucose concentrations that occur after ingestion of a
carbohydrate-rich meal.

Hexose
transporters also
have fairly
distinctive
patterns of
expression among
tissues. Most cells
express more than one hexose transporters.

Consider for example, small intestinal epithelial cell, which

must absorb glucose, galactose and fructose from the intestinal
lumen, then export those sugars into blood. This cell has at least
three hexose transporters. The apical membrane contains the
sodium-glucose cotransporter SGLUT1, which allows the cell to
take up glucose and galactose by cotransport with sodium, and
GLUT5, which mediates absorption of fructose. On the
basolateral plasma membranes is GLUT2, which allows diffusion
of all three of these hexoses out of the cell into extracellular
fluid and ultimately, into blood.

The hexose transporters are large

integral membrane proteins. Based on
the deduced amino acid sequences of
their cloned cDNAs, they have similar
structures, consisting of 12
membrane-spanning regions with
cytoplasmic C-terminal and N-terminal tails. Also, they all
appear to be glycosylated on one of the extracellular loops.

Transport of sugars across membranes appears to result from a

series of confirmational changes which "flips" the transporter
between alternate states with the substrate binding site either
facing the extracellular or cytoplasmic side of the membrane.
Transport in either direction is thus possible, depending on
relative substrate concentrations on either side of the
membrane.

Salient characteristics of the glucose transporters are

summarized in the table below.

Major Sites
Transporter Characteristics
of Expression
 Brain,
Transports glucose (high
erythrocyte,
affinity) and galactose, not
GLUT-1 endothelial
fructose. Expressed in many
cells, fetal
cells.
tissues

Tranports glucose, galactose

Liver,
and fructose. A low affinity,
pancreatic beta
high capacity glucose
GLUT-2 cell, small
transporter; serves as a
intestine,
"glucose sensor" in
kidney.
pancreatic beta cells.

Transports glucose (high

affinity) and galactose, not
Brain, placenta
GLUT-3 fructose. The primary
and testes
glucose transporter for
neurons.

Skeletal and The insulin-responsive

GLUT-4 cardiac muscle, glucose transporter. High
adipocytes affinity for glucose.

Transports fructose, but not

Small intestine, glucose or galactose. Present
GLUT-5
sperm also in brain, kidney,
adipocytes and muscle
MECHANISM OF GLUCOSE TRANSPORT
Glucose is absorbed through the intestine by a transepithelial
transport system initiated at the apical membrane by the
cotransporter SGLT-1; intracellular glucose is then assumed to
diffuse across the basolateral membrane through GLUT2. Here,
we evaluated the impact of GLUT2 gene inactivation on this
transepithelial transport process. We report that the kinetics of
transepithelial glucose transport, as assessed in oral glucose
tolerance tests, was identical in the presence or absence of
GLUT2; that the transport was transcellular because it could be
inhibited by the SGLT-1 inhibitor phlorizin, and that it could not
be explained by overexpression of another known glucose
transporter. By using an isolated intestine perfusion system, we
demonstrated that the rate of transepithelial transport was
similar in control and GLUT2−/− intestine and that it was
increased to the same extent by cAMP in both situations.
However, in the absence, but not in the presence, of GLUT2, the
transport was inhibited dose-dependently by the glucose-6-
phosphate translocase inhibitor S4048. Furthermore, whereas
transport of [14C]glucose proceeded with the same kinetics in
control and GLUT2−/− intestine, [14C]3-O-methylglucose was
transported in intestine of control but not of mutant mice.
Together our data demonstrate the existence of a
transepithelial glucose transport system in GLUT2 −/− intestine
that requires glucose phosphorylation and transfer of glucose-6-
phosphate into the endoplasmic reticulum. Glucose may then
be released out of the cells by a membrane traffic-based
pathway similar to the one we previously described in GLUT2-
null hepatocytes. After intake and digestion, carbohydrates are
absorbed in the small intestine of mammals by a two-step
transport system. In the first step, glucose is concentrated in
the cells by a mechanism catalyzed by the apically located
sodium-dependent glucose transporter SGLT-1. This symporter
uses the electrochemical gradient of two sodium ions to
transport one glucose molecule. The second step is the release
of the intracellular glucose into the interstitial space by a
mechanism thought to occur by facilitated diffusion via the
glucose transporter GLUT2 located in the basolateral
membrane.

It is generally believed that SGLT-1 is the only translocator for

entry of glucose into the enterocytes. The most forward
evidence comes from patients suffering from glucose/galactose
malabsorption, a potentially fatal condition, which is caused by
inactivating mutations in SGLT-1. In contrast, the contribution of
GLUT2 to the second step of transepithelial glucose transport
rests mainly on its immunolocalization to the basolateral
membrane domain and on studies of glucose transport in
vesicles derived from these membranes, which showed
properties compatible with the presence of GLUT2. Thus,
whereas GLUT2 may contribute to the second step of
transepithelial glucose transport, separate mechanisms also
may exist.
Defects
Defective transport not only results in malabsorption, but also
accumulation of metabolic intermediates, often with toxic
consequences. Other defects play a role in energy coupling
(mitochondrial and peroxisomal disorders) or cell volume
regulation. Control of water flow is important for solubilization
and distribution of metabolites, body temperature regulation
through sweating, and waste secretion. Many bacterial toxins
negatively affect a cell's ability to regulate water and ion
absorption and retention causing loss of water (diarrhea) or the
accumulation of metabolites.

Glucose Galactose Malabsorption is a genetic disorder caused

by a defect in glucose and galactose transport across the
intestinal brush border. Normally, lactose in milk is broken
down into glucose and galactose by lactase, an ectoenzyme on
the brush border, and the hexoses are transported into the cell
by the Na+-glucose cotransporter SGLT1. The mutations causing
the defect in sugar transport have been identified in patients
from 33 kindreds, and functional studies have established how
these mutations cause the disease.

Defects in transport function or its regulation can severally

affect homeostasis and causes excessive water retention or loss.
Diarrhea is one of the most common disease and caused by
many different factors related to neuronal, immunological,
hormonal regulation, or lumenal content, particularly bacterial
toxins. Well known (but not necessarily well understood in
terms of mechanism) causes of diarrhea are Cholera,
Inflammatory Bowel Syndrome (Crohn's Disease), Celiac disease
(also known as celiac sprue and gluten-sensitive enteropathy),
lactose intolerance, and AIDS related diarrhea. While cholera
and AIDS related diarrhea are related to bacterial infections and
release of toxins, Crohn's and celiac disease are the result of
damaged mucosal epithelia due to overractive immune
responses and altered cell adhesion mechanisms. Lactose
intolerance and the related glucose-galactose malabsorption
result in accumulation of mono- and disaccharides in the
intestinal lumen and exert osmotic pressure on the epithelia
resulting in the loss of water. Glucose-galactose malabsorption
is well understood at a mechanistic level. Here, the apical Na-
coupled transporter SGLT1 is defective and cannot be
transported to the cell surface. In a functional system, glucose is
transported into the enterocyte along with a sodium ion.
Glucose then diffuses into the portal blood via the glucose-
specific transporter GLUT2. Sodium leaves the enterocyte via
the Na/K pump on the basal membrane and potassium is
recycled through a K-channel. This energy driven uptake of
glucose and sodium serves as a prototype mechanism of
absorption and secretion.

Many transport defects affecting the gut also affect other

organs that rely on epithelial absorption and secretion
mechanism. This includes the kidney, sweat glands, and lungs.
Renal glucosuria is a condition with enhanced glucose in the
urine. The cause is defective reabsorption of glucose from
filtered blood serum fluid in the nephrons of the kidney. The
defect is related to the sodium coupled glucose transporter.
Patients with glucosuria have no intestinal glucose uptake
defect, indicating that a different transporter, SGLT2, is
expressed in the kidney epithelia.
Inhibitors
Inhibitors of glucose transport have been especially useful for
detecting and distinguishing between facilitated and secondary
active glucose transport. Much like enzymes there are both
competitive and non-competitive inhibitors of glucose
transport. Review the concepts of competitive and non-
competetive enzyme inhibition in your text. Cytochalasin B is a
non-competitive inhibitor of glucose uptake by GLUT proteins,
but not by SGLT proteins. Likewise, Phloretin (a mold
metabolite) inhibits GLUT protein transport, while Phloridzin (a
different plant glycoside) inhibits Na+ coupled transport
(Hediger and Rhoads, 1994). Interestingly, both phloretin and
phloridzin act through a competitive inhibition mechanism.

Hormones regulate the expression and/or the localization of

some glucose transporter proteins. In some cells hormones are
able to stimulate glucose transport. This may be by direct
mobilization of the transport protein to the membrane as in the
case of insulin regulated mobilization of GLUT4 protein (Karp,
2005), or by increases in transcription and/or translational from
GLUT genes (Rodolfo et al., 2003).

A small amount of glucose transport can occur when protein-

mediated transport is inhibited by competing sugars or by other
means. This is seen as a low baseline level of glucose transport
in experiments that monitor transport by radioactively labeled
glucose. This transport is minimal, seemingly non-regulatable
and generally not considered to be physiologically important.

References
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3.Thorens B: Facilitated glucose transporters in epithelial cells.

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