7141665

The Role of Odorant-Binding Proteins

An odorant-binding protein (OBP) is a low molecular weight, soluble

protein which can be found in high concentrations in the nasal mucus of
vertebrates and the lymph of sensilla in insects. Although the physiological role
of these proteins has not yet been clarified, they are strongly believed to have a
role in olfactory reception and possibly even perception, although no direct
evidence exists for the latter in vertebrates. OBPs have a main affinity for odours
and pheromones, and several members have been identified and sequenced.
Several types of OBP are expressed in the nasal epithelium, and their sequence
shows similarity to a large family of small hydrophobic molecules know as the
lipocalins, which have a role in retinoid and pheromone binding[Pelosi, 1994].
To understand the function of an OBP we must first understand what is
meant by an odour. Unfortunately this doesn’t aid scientists much as odours
aren’t a specific class of molecules but rather a molecule which can stimulate
olfactory neurons. For animals who breathe air this means the molecule must be
volatile enough to reach the olfactory receptors in the nasal epithelium, which
limits these molecules to low molecular weight and weak intermolecular forces.
However this is not the case in aquatic creatures as polar compounds easily
dissolve in aqueous solutions due to their charged nature, making them
accessible odourants in liquid environments. The strength of an odour is
measured by its olfactory threshold, i.e. the lowest concentration at which a
significant sample of a population can perceive the odour. Strong odours are the
most useful probes in research settings as it’s believed that these would make
the strongest interactions with olfactory receptors. This rather weak classification
of an odour (at least from a physiological view point) makes it hard to
understand the true function of an OBP as there are such a large diversity of
odours that they are required to bind to.

A closer looks at the biochemical properties of OBPs can help clarify their
function. As previously stated, they are soluble proteins, a necessity since they
are located in aqueous solutions such as mucus and lymph. Odours are
hydrophobic, and so OBPs have been suggested to aid solubility of these
molecules to amplify the signal. They are mostly homodimers, i.e. they are made
up of two identical subunits [Fesenko, 1987], however some OBPs do exist as
monomeric species. High concentrations of OBPs can be found in the olfactory
epithelium and sensillar lymph[Klein, 1987], which is one of the few ways in
which we can classify a protein as an OBP. Their rather weak protein-ligand
interactions with odours means they may have a temporary association with
odours, which would suggest a role in transportation of odours, and also a broad
specificity to odour molecules, meaning one OBP can bind to several different
odours. This indicated a possible function as a mechanism for odour perception,
whereby a few broadly specific OBPs can create a code for the recognition of
many odours.

OBPs fall under the lipocalin family of proteins, and have sequence
similarity to the proteins which fall under this category. These proteins are
generally involved in the transportation of hydrophobic molecules in solution.
The similarity in structure and sequence would indicate a highly probability that
OBPs perform a similar function to other lipocalin proteins. OBPs do differ from
lipocalins in at least one respect, that being they have broad specificity, meaning
they can bind to many different ligands. Most lipocalins on the other hand have a
much higher affinity and selectivity. This aids the job of determining OBP
7141665

function by distinguishing it from other lipocalins, but also hinders it at the same
time due to the huge diversity in molecules in which they can bind to.

A major problem with determining OBP function is their low levels of

conservation in the genome, sharing approximately 30% of their amino acids.
This makes it hard to classify a protein as an OBP as it removes the ability to
group proteins by their sequence similarity. The diversity of OBPs is also evident
in their tissue localisation, as even though they are found only in the nasal
epithelium and sensillar lymph, they are produced by different glands in different
areas. Not only this but they are synthesised in regions which are separate to the
ciliated olfactory epithelium. This however does not mean they are not a part of
the olfactory detection mechanism as several proteins in other systems are
produced in areas which are not where their function is aimed to take place.
Isoelectric points have also been found to differ slightly, ranging from 4 to 5. The
diversity of OBPs and their supposed target molecules does pose problems when
it comes to studying their physiological role, however the discovery of several
different types of OBP in the same species does further the possibility that their
role lies in the discrimination of odours.

The difficultly found when trying to define and class OBPs reflects the
numerous problems found when trying to determine their role. Ideally if we could
classify a protein as an OBP solely on their ability to bind to odours, this would be
a much easier proposition, and would help in distinguishing these proteins from
other similar ones in the lipocalin family. However, the diversity in odours and
their biochemical properties make this impossible, as most odours have other
roles to play than to just be detected as a “smell”. A weak but useful guideline
for OBP specificity was determined form binding experiments. For example,
many odorants of varying structure show at least some affinity to OBP, and none
of them show very strong affinity. Compounds don’t necessarily have the same
ability to bind to OBP just because they’re in the same chemical class, it tends to
depend more on their level of hydrophobic interaction, and therefore alkyl chains
of longer length have a lower binding constant. Structure of odours does show
some patterns. Round shaped odours tend to show no significant binding,
whereas planar molecules (e.g. ones with carbon rings) tend to show relatively
strong interaction. Long, flexible molecules such as alcohols with long carbon
chains also show a relatively strong interaction with OBPs, but this may be
because they can bend into a planar or ring shape similar to those of the other
high affinity ligands[Bucci, 2006]. The fact that different odours bind with
different degrees of strength shows to some extent a degree of discrimination,
which strengthens the odour perception hypothesis. [Pelosi, 1994]

In an experiment involving odours of similar structure and their ability to

bind to bovine OBP, ‘green’ smells such as thiazoles had a much stronger affinity
to the OBP. This makes sense as a cow’s main diet consists of grasses and other
vegetation. This shows that OBPs have the potential to fall under selection
pressure, even though they are very poorly conserved[Topazzini, 1985].
Pyrazines followed a similar pattern, showing a strong degree of binding for
porcine and bovine OBPs. [Pelosi, 1994][Baldaccini, 1986] [Pevsner, 1985]

When considering the three dimensional structure of most OBPs, the

subunits show structural similarity to the lipocalins. Bovine OBP is folded into 8
anti-parralel B-pleated sheets, with a single a-helical region. This a-helix
supposedly gives the molecule flexibility, making it easy to bind with some
affinity to a wide range of molecules with a wide range of structures. On its own,
7141665

the subunit wouldn’t have a large enough region for binding to odours, however,
when bound to its counterpart they create a large hydrophobic region where the
hydrophobic odours can sit, allowing them to enter into aqueous environments
like the nasal mucus.

Insects OBPs are found in the sensillar lymph, but they show very little
similarity to vertebrate OBPs. They are more highly conserved though, and the
organ which they exist in, the sensilla, has a very specific function in olfaction,
making it much easier to determine their main function. Like vertebrate OBPs
they exist in high concentrations in specific areas. Even though they do differ
from species to species they have common characteristics to vertebrate OBPs
(small molecular weight, low isoelectric points and high abundance) showing
their place as the invertebrate version of OBPs. [De Kramer, 1987]. This also
means that their function may well be very similar too; meaning vertebrate OBPs
could share the same function as insect OBPs at the peripheral level[Getchell,
1984]. Three types of insect OBPs exist; general odourant-binding proteins
(GOBPs), pheromone-binding proteins (PBPs) and chemosensory proteins (CSPs)
although the latter could be considered entirely separate [Pelosi, 2006]. GOBPs
are have a broader specificity than PBPs, and therefore may show greater
resemblance in function to vertebrate OBPs. This is again due to weak non-
specific protein-ligand interaction[Lescop, 2009]. Two genes for OBPs in
Drosophila melanogaster were studied by Harada et al. (2008). Obp57d and
Obp57e were found to be involved in plant preference due to a be havioural
response to acids. Homozygotes showed very different responses to the acid, but
interestingly heterozygotes showed a significantly different response compared
to the wild-type, which suggest a gene dosage effect on the behaviour of D.
melanogaster. These genes were highly associated with the determination of
reproductive sights, rather than locating food. OBPs in this case show an
important role in ecological adaptation.

When taking into account all this information, it’s easy to see why no clear
function for OBPs has been obtained. This hasn’t stopped scientists from
speculating as to what the function of these proteins could be. The main
question which appears to come to mind is are OBPs involved in odour
perception/discrimination, i.e. are OBPs responsible for telling odours apart
[Dear, 1991]. The fact that they are only produced in the nasal cavity and
sensilla, both organs associated with olfaction, makes it fairly refutable to say
that they are involved in the process of detecting odours. Sensilla being much
specialised in their function, contain high concentrations of insect OBPs. Their
similarity in characteristics implies a similar function for vertebrate OBPs. Several
hypotheses have been suggested for OBPs ability to bind to odours. One
hypothesis suggests that OBPs transport odours to the receptor membrane,
which would aid detection by increasing odour solubility in the mucus layer,
effectively amplifying the signal and increasing the ability to detect odours
[Pevsner, 1990]. Another hypothesis suggests a similar function, but instead of
transporting odours to the membrane, they remove them from the receptor to
allow other molecules to bind [Boudjelal, 1996].) A hypothesis which is quite
different to the first two suggests that OBPs filter and buffer odours by trapping
them. This means that their function would only be necessary when high
concentrations of odour enter the nose, which could cause inactivation of the
receptor for a long period of time. The existence of more than one OBP in a
single organism would make it possible to discriminate between odours, and
their broad specificity would also help cater to this ability. Although it’s been
shown that odours can directly activate odour receptors, it goes without question
7141665

that OBPs significantly enhance this process, and although an exact function has
not yet been proven, it’s more than reasonable to think there’s a good chance
they could help create a code which enables the brain to differentiate between
these stimuli. Their role in olfaction is not limited to the acquisition of food either,
as seen by the Obp57d and Obp57e genes, but can also affect behavioural
ecology in relation to reproducing. Hopefully in future studies, a way to view
OBPs function in vivo will be developed, to help try and solve once and for all
their true role in life.

Bibliography
Baldaccini, N. G. (1986). Occurence of a pyrazine binding protein in the nasal
mucosa of some vertebrates. Compar. Biocehm. Physiol (84B), 249-253.

An interesting study involving the binding of pyrazine to OBPs found in cow and
pig. Showed the evolutionary advantages to OBP differentiation and specificity
based on environment and nutrition.

Boudjelal, M. S. (1996). Membrane receptor for odour-binding proteins.

Biochemistry Journal (317), 23-27.

A summary of odour-binding proteins and their potential for their role in

transporting odours to/from the receptor membrane.

Bucci, B. K. (2006). Effect of n-alcohols on the structure and stability of the

Drosophila odorant binding protein LUSH. Biochemistry (45), 1693-1701.

A look at alcohol binding to the LUSH OBP found in fruit flies, and how chain
length can affect affinity.

De Kramer, J. J. (1987). The neurobiology of pheromone reception. In J. J. De

Kramer, Pheromone Biochemistry (pp. 433-472). Orlando: Academic Press.

A study which looks at pheromone communication in flies, and particularly the

role of PBPs.

Dear, T. B. (1991). Novel genes for potential ligand-binding proteins in

subregions of the olfactory mucosa. Embo. J. , 10, 2813-2819.

An in depth study of OBPs in vertebrates. Focusing on their specificity of where

they are produced and their potential role as differentiation of odours.

Fesenko, E. N. (1987). The subunits of specific odor-binding glycoproteins form

rat olfactory epithelium. FEBS. Lett. (219), 224-226.

Getchell, T. M. (1984). Perireceptor and receptor events in vertebrate olfaction.

Prog. Neurobiol. (23), 317-345.

Harada, E. H. (2008). Behavioural analyses of mutants for two odorant-binding

protein genes. Genes Genet. Syst. (83), 257-264.
7141665

Klein, U. (1987). Sensillum lymph proteins from antennal olfactory hairs of the
moth Antheraea polyphemus (Saturnidae). Insect Biochem. (17), 1193-1204.

Lescop, E. B. (2009). Structural basis of the braod specificity of a general

odorant-binding protein from honeybee. Biochemistry (48), 2431-2441.

Pelosi, P. (1994). Odorant-binding proteins. Critical REviews in Biochemistry and

Molecular Biology , 3 (29), 199-228.

Pelosi, P. Z.-J. (2006). Soluble proteins in insect chemical communication. Cell.

Mol. Life Sci. , 63, 1658-1676.

Pevsner, J. S. (1990). Odorant-binding protein: odorant transport function in the

vertebrate nasal epithelium. Chem. Senses (15), 217-222.

Pevsner, J. T. (1985). Isolation and characterization of an olfactory receptor

protein for odorant pyrazines. Proc. Natl. Acad. Sci. U.S.A. , 82, 3050-3054.

Topazzini, A. P. (1985). Specificity of pyrazine binding protein from cow olfactory

mucosa. Chem. Senses (10), 45-49.