PERGAMON

The International Journal of Biochemistry & Cell Biology 30 (1998) 711

Biochemistry
&
Cell Biology

Molecules in focus

The uncoupling protein, thermogenin

Andreu Palou, Catalina Pico, M. Luisa Bonet, Paula Oliver
Laboratori de Biologia Molecular, Nutricio i Biotecnologia. Departament de Biologia Fonamental i Ciencies de la Salut. Universitat
de les Illes Balears. Ctra. Valldemossa km 7.5, Palma de Mallorca07071, Spain
Received 3 March 1997

Abstract
The uncoupling protein (UCP) or thermogenin is a 33 kDa inner-membrane mitochondrial protein exclusive to
brown adipocytes in mammals that functions as a proton transporter, allowing the dissipation as heat of the proton
gradient generated by the respiratory chain and thereby uncoupling oxidative phosphorylation. Thermogenesis (heat
production) in brown adipose tissue, which is activated in response to cold exposure or chronic overeating, depends
largely on UCP activity. Norepinephrine, released from sympathetic terminals and acting via b-adrenoceptors and
cAMP, is the main positive regulator of both UCP synthesis and activity. Brown fat thermogenesis plays a critical
role in thermoregulation and in overall energy balance, at least in rodents. Manipulation of thermogenesis, whether
through UCP or through analogous uncoupling proteins, could be an eective strategy against obesity. # 1998
Elsevier Science Ltd. All rights reserved.
Key words: Uncoupling protein; Thermogenin; UCP; UCP2; Obesity; Thermogenesis

1. Introduction
White and brown adipose tissues (WAT and
BAT) serve important opposite functions in overall energy balance. While WAT is specialized in
energy storage in the form of triacylglycerols,
BAT functions to dissipate energy in the form of
heat (thermogenesis). BAT is especially abundant
Abbreviations: UCP, uncoupling protein, BAT, brown adipose
tissue,WAT, white adipose tissue, SNS, sympathetic nervous system, NE, norepinephrine, T3, triiodothyronine, PKA, protein
kinase A, CREB, cAMP-response-element-binding protein,
RXRa, retinoid X receptor a, RARa, retinoic acid receptor a.

in small mammals, but it is present in the young

of most mammalian species, even in humans.
Brown adipocytes are smaller than white adipocytes, contain less lipid that is stored in multilocular rather than unilocular droplets, and possess
abundant mitochondria with a unique feature:
the presence in their inner membrane of the
uncoupling protein (UCP). BAT thermogenesis
depends largely on the activity of this protein.
BAT growth and thermogenic function are
under the regulation of the sympathetic nervous
system (SNS), which densely innervates the tissue
(reviewed in Himms-Hagen, 1990). They are
induced under physiological conditions associated

0003-9969/98/$19.00 # 1998 Elsevier Science Ltd. All rights reserved.

PII: S 1 3 5 7 - 2 7 2 5 ( 9 7 ) 0 0 0 6 5 - 4

A. Palou et al. / The International Journal of Biochemistry & Cell Biology 30 (1998) 711

with a chronic increase in the sympathetic stimulation to BAT, with enhanced norepinephrine
(NE) release to the tissue, such as cold exposure
or chronic overfeeding, in an adaptative response
that tends to resist cold and body weight gain,
respectively. Conversely, a chronic decrease in
SNS activity to BAT, as occurs during cold-deacclimation and fasting, induces atrophy and
decreases thermogenesis.
2. Structure
UCP is a 33 kDa monomer of 306 amino
acids; the functional unit may be an homodimer.
Its amino acid sequence is highly homologous
among species, suggesting its biological importance; it is also highly homologous to that of several ubiquitous mitochondrial inner-membrane
carriers, including the ADP/ATP carrier, the
phosphate carrier and the oxoglutarate carrier.
From the sequence data, it has been suggested
that UCP consists of three membrane-spanning
``Us'', each of about 100 amino acids (Fig. 1)
and encoded by two exons separated by one
intron in the UCP gene, which is nuclear and
single-copy. The overall protein structure consists
of about 50% a-helix, 30% b-structure, 15% bturns and 7% random (reviewed in Nedergaard
and Cannon, 1992).

,,
,
,,
,
,,
,
,,
,,,
,,
,,
Intermembrane space

N-ter.
1

11

33

Inner
mitochondrial
membrane

89

111 188

67 133

C-ter.

210 282

166 232

306

260

Mitochondrial matrix

Fig. 1. Tripartite structural model of the uncoupling protein

(adapted from Nedergaard and Cannon, 1992 and Bouillaud
et al., 1994). The protein (306 amino acids) has been proposed
to constitute three symmetrical membrane-spanning ``Us'',
each comprising about 100 amino acids, and to contain six
amphiphilic transmembrane a-helices (represented here by
boxes). Numbers correspond to the position in the primary
structure of the amino acid residues that delimitate the transmembrane domains.

3. Synthesis and degradation

NE has a main role on brown adipocytes, promoting thermogenesis itself (UCP activation),
dierentiation (UCP synthesis) and cell division
(reviewed in Cannon et al., 1996). The major
adrenoceptor mediating the eects of NE in
mature BAT cells is b3-adrenoceptor, which is
mainly expressed in white and brown adipocytes:
a1-, b1- and b2-adrenoceptors are also involved,
though to a lesser extent. b-Adrenoceptors are
coupled via stimulatory G-proteins to adenylyl
cyclase: their stimulation by NE leads to an
increase of cAMP that in turn stimulates protein
kinase A (PKA) activity. Key downstream targets
of PKA include hormone-sensitive lipase, which
once phosphorylated actively breaks down triacylglycerol stores providing fatty acids to activate
thermogenesis (see below), and CREB (cAMP-response-element-binding protein), which stimulates
transcription from the UCP gene. In primary
brown adipocytes dierentiated in culture, NE
also stabilizes UCP mRNA (Pico et al., 1994).
The UCP gene is mainly regulated at the transcriptional level (reviewed in Susulic and Lowell,
1995). Besides NE, another activator of UCP
transcription of physiological relevance is triiodothyronine (T3), which acts in connection with
NE, since NE stimulation markedly increases
thyroxine deiodinase activity of brown adipocytes, thus allowing high intracellular generation
of T3 from thyroxine. Retinoic acid, the natural
active form of vitamin A, has been shown to
stimulate UCP gene transcription through a
mechanism independent from the adrenergic
pathway, and to induce UCP appearance both in
cultured brown adipocytes and in BAT of intact
mice (Larose et al., 1996; Puigserver et al., 1996;
and references therein). The DNA sequences in
the UCP gene promoter that confer brown fatspecic expression and responsiveness to cAMP,
T3 and retinoic acid have been identied; for the
latter three, the likely trans-acting factors are
CREB, T3 receptor-RXRa heterodimers, and
RAR-RXRa heterodimers, respectively.
Concerning UCP degradation, there is evidence
for the existence of dierent pools of UCP of
dierent half-life that are probably related to

A. Palou et al. / The International Journal of Biochemistry & Cell Biology 30 (1998) 711

dierent mitochondrial subpopulations, and it

has been suggested that a post-translational regulatory mechanism exists which ensures the rapid
degradation of newly synthesized molecules when
the physiological stimulation ceases (Puigserver
et al., 1992; Bonet et al., 1995).
4. Biological function
UCP acts as a highly regulated proton transporter over the mitochondrial inner membrane.
Its proton transport activity is inhibited by the
binding of purine nucleotides (mainly ATP,
under the cellular conditions) to a site on UCP
itself, and activated by fatty acids. When stimulated, UCP mediates the passive re-entry of protons into the mitochondrial matrix: this produces
heat and a concomitant decrease in the ATP
yield of oxidative phosphorylation (Fig. 2). As a
consequence, oxidative metabolism in the brown
adipocyte proceeds at a maximum rate, enabling
its mitochondria to produce more heat.
Two models have been proposed to explain
fatty acid-mediated stimulation of UCP (reviewed
in Susulic and Lowell, 1995). In the rst, UCP is
seen as a proton transporter, and fatty acids
H+
H+
ATP-synthase

H+ H+
+
H+ H +
H
H+ H+

UCP
+
H+ H+
+
H
H

+
H+ H H+
+
H+ H+ H+ + H+
H
H
H+ H+ H+

Respiratory chain

H+

H+

H+

O2
H2O

H+

ATP
synthesis

Heat
protection

Fuel
oxidation

Fig. 2. The three conceptual elements involved in the function

of the uncoupling protein: fuel oxidation, energy dissipation
and energy conservation. During fuel oxidation protons are
released and pumped out from the mitochondria by the respiratory chain. In stimulated brown adipocyte mitochondria,
they may re-enter either through the ATP-synthase (as in normal mitochondria) or through the UCP. In the latter case, the
energy stored in the proton electrochemical gradient is dissipated as heat (thermogenesis) instead of being conserved as
ATP.

serve as allosteric activators of proton transport.

In the second, UCP is seen as a fatty acid anion
transporter: fatty acid anions themselves, which
are membrane impermeable, are transported out
of the mitochondria by UCP, become protonated
in the cytoplasm and re-enter the mitochondria
in this membrane-permeable state, thus acting as
cycling protonophores.
Fatty acids not only interact directly with
UCP, but also are the main substrate oxidized by
brown fat cells mitochondria. Therefore, when
lipolysis of endogenous triacylglycerol stores is
stimulated by NE, fatty acids provide both the
intracellular signal and the fuel for the activation
of thermogenesis. During chronic adrenergic
stimulation, BAT obtains fatty acids from plasma
lipoproteins via its lipoprotein lipase, the expression of which is stimulated by NE.

5. Role in energy balance and obesity; therapeutic

implications
There is strong evidence that UCP-mediated
BAT thermogenesis represents a chapter of
energy expenditure of critical importance in overall energy balance, at least in rodents. A decient
BAT thermogenesis is found in practically all
animal models of obesity (reviewed in HimmsHagen, 1990); BAT-ablated transgenic mice
develop obesity (reviewed in Susulic and Lowell,
1995); genetic manipulations that enhance the
levels of UCP prevent genetic and diet-induced
obesity in mice (Kopecky et al., 1995; Cummings
et al., 1996); and it has been suggested that the
increase in energy expenditure brought about by
leptin, the product of the ob gene, involves activation of NE-stimulated BAT thermogenesis (see
Fig. 3).
Although the physiological signicance of BAT
in humans is less clear, the functionality of the
b3-UCP thermogenic system is becoming evident.
BAT, albeit diuse and at levels greatly reduced
compared with neonates, is present in adults and
retains capacity of expansion. The human b3adrenoceptor appears to be functional (Strosberg
and Pietri-Rouxel, 1996), and there is a corre-

10

A. Palou et al. / The International Journal of Biochemistry & Cell Biology 30 (1998) 711

Fig. 3. Heat production in BAT in the framework of the overall body weight control system. The scheme shows how obesity can
result from a lower activity of the leptinneuropeptide Y pathway, a defect of a putative leptinmelanocortin pathway or a defect
in the b3-UCP thermogenic system. Norepinephrine, insulin and cytokines could connect the dierent elements of this body weight
control system, as shown in the scheme. Genetic and experimentally-induced alterations aecting the dierent pink-coloured components of the scheme have been shown to be associated with body weight disregulation. Up to now, statistical association of
genes and obesity has been shown in human populations only for the b3-adrenoceptor and the UCP genes. CM, chylomicrons;
CNS, central nervous system; CREB, cAMP-response-element-binding protein; FA, fatty acids; HSL, hormone sensitive lipase;
LPL, lipoprotein lipase; MSH, melanocyte-stimulating hormone; NPY, neuropeptide Y, triacylgycerols; VLDL, very low-density
lipoproteins.

lation in human populations between the expression of certain genetic variants of the b3adrenoceptor and the UCP genes, and resistance
to weight loss and/or development of obesity
over years (see Clement et al., 1995, 1996).
Taken together, these data favour the idea that
BAT, or BAT-like systems, also play a role in
human energy balance, and have raised a considerable interest in it as the target of anti-obesity
drugs/treatments aimed at enhancing thermogenesis (as, for example, b3-agonists).

Most relevant to the obesity problem is the

existence of other thermogenic UCP-like molecules, such as the recently discovered UCP2
(Fleury et al., 1997), an uncoupling protein that
shares a considerable sequence homology with
UCP (from now on UCP1) and of apparent analogous function, but with a very dierent regulation, since it does not respond either to cold or
to b3-agonists. UCP2 is widely expressed in both
human and rodent tissues, maps to chromosomic
regions linked to obesity and hyperinsulinemia

A. Palou et al. / The International Journal of Biochemistry & Cell Biology 30 (1998) 711

and is upregulated in WAT of mice fed a highfat diet. Interestingly, UCP2 is induced in BAT
of ucp1 knockout mice, which are cold-sensitive
but do not develop obesity (Enerback et al.,
1997). Whether UCP2 is normally involved in
burning away extra calories, the molecular mechanisms that regulate its uncoupling function, and
the possible existence of other members of a
family of uncoupling proteins, are the next questions to be addressed. The answers, properly integrated into what we know about the complex
and not yet fully understood apparatus controlling mammalian body weight, should help in
designing new strategies against obesity and its
complications.
Acknowledgements
This work was funded by DGICYT of the Spanish
Government (grants PB-92-0748-C04-01 and PB94-1178) and
by
the
European
Comission
(Contract
No.
ERBCHRX#CT940490).

References
Bonet, M. L., Serra, F., Matamala, J. C., Garc| a-Palmer, F.
J. and Palou, A. (1995) Selective loss of the uncoupling protein of light versus heavy mitochondria of brown adipocytes
after a decrease in noradrenergic stimulation in vivo and in
vitro. Biochem. J. 311, 327331.
Bouillaud, F., Arechaga, I., Petit, P. X., Raimbault, S., LeviMeyrueis, C., Casteilla, L., Laurent, M., Rial, E. and
Ricquier, D. (1994) A sequence related to a DNA recognition element is essential for the inhibition by nucleotides
of proton transport through the mitochondrial uncoupling
protein. EMBO J. 13, 19901997.
Cannon, B., Jacobsson, A., Rehnmark, S. and Nedergaard, J.
(1996) Signal transduction in brown adiposse tissue recruitment: noradrenaline and beyond. Int. J. Obesity 20, S36
S42.
Clement, K., Ruiz, J., Cassard-Doulcier, A. M., Bouillaud,
F., Ricquier, D., Basdevant, A., Guy-Grand, B. and
Froguel, P. (1996) Additive eect of A 4 G(-3826) variant
of the uncoupling protein gene and the Trp64Arg mutation
of the b3-adrenergic receptor gene on weight gain in morbid obesity. Int. J. Obesity 20, 10621066.
Clement, K., Vaisse, C., Manning, B. S. T., Basdevant, A.,
Guy-Grand, B., Ruiz, J., Silver, K. D., Shuldiner, A. R.,
Froguel, P. and Strosberg, D. (1995) Genetic variation in

11

the b3-adrenergic receptor and increased capacity to gain

weight in patients with morbid obesity. N. Engl. J. Med.
333, 343347.
Cummings, D. E., Brandon, E. P., Planas, J. V., Motamed,
K., Idzerda, R. L. and McKnight, G. S. (1996) Genetically
lean mice result from targeted disruption of the RIIb subunit of protein kinase A. Nature 382, 622626.
Enerback, S., Jacobsson, A., Simpson, E. M., Guerra, C.,
Yamashita, H., Harper, M. E. and Kozak, L. P. (1997)
Mice lacking mitochondrial uncoupling protein are coldsensitive but not obese. Nature 387, 9094.
Fleury, C., Neverova, M., Collins, S., Rainbault, S.,
Champigny, O., Levi-Meyrueis, C., Bouillard, F., Seldin,
M. F., Surwit, R. S., Ricquier, D. and Warden, C. H.
(1997) Uncoupling protein-2: a novel gene linked to obesity
and hyperinsulinemia. Nature Genet. 15, 269272.
Himms-Hagen, J. (1990) Brown adipose tissue thermogenesis:
interdisciplinary studies. FASEB J. 4, 28902898.
Kopecky, J., Clarke, G., Enerback, S., Spiegelman, B. and
Kozak, L. P. (1995) Expression of the mitochondrial
uncoupling protein gene from the aP2 promoter prevents
genetic obesity. J. Clin. Invest. 96, 29142923.
Larose, M., Cassard-Doulcier, A. M., Fleury, C., Serra, F.,
Champigny, O., Bouillaud, F. and Ricquier, D. (1996)
Essential cis-acting elements in rat uncoupling protein gene
are in an enhancer containing a complex retinoic acid response domain. J. Biol. Chem. 271, 3153331542.
Nedergaard J. and Cannon B. (1992) The uncoupling protein
thermogenin and mitochondrial thermogenesis. In New
Comprehensive Biochemistry, Molecular Mechanisms in
Bioenergetics (Edited by Ernster L.), Vol. 23, pp. 385420.
Elsevier, Amsterdam.
Pico, C., Herron, D., Palou, A., Jacobsson, A., Cannon, B.
and Nedergaard, J. (1994) Stabilization of the mRNA for
the uncoupling protein thermogenin by transcriptional/
translational blockade and by noradrenlaine in brown adipocytes dierentiated in culture: a degradation factor
induced by cessation of stimulation?. Biochem. J. 302, 81
86.
Puigserver, P., Herron, D., Gianotti, M., Palou, A., Cannon,
B. and Nedergaard, J. (1992) Induction and degradation of
the uncoupling protein thermogenin in brown adipocytes in
vitro and in vivo. Biochem. J. 284, 393398.
Puigserver, P., Vazquez, F., Bonet, M. L., Pico, C. and Palou,
A. (1996) In vitro and in vivo induction of brown adipocyte
uncoupling protein (thermogenin) by retinoic acid.
Biochem. J. 317, 827833.
Strosberg, A. D. and Pietri-Rouxel, F. (1996) Function and
regulation of the b3-adrenoceptor. Trends Pharmacol. Sci.
17, 373381.
Susulic, V. S. and Lowell, B. B. (1995) Brown adipose tissue
and the regulation of body fat stores. Curr. Opin.
Endocrinol. Diabetes 3, 4450.