Abstract. Objective and Design: Investigation of the prin-
ciples of ligand-receptor interaction in histamine receptors
can help to provide a solid foundation for structure-based
drug design. Stable isotope labelling of the ligand ‘Hista-
mine’ has been performed and 1D
13
C CP MAS and 2D
Radio Frequency Dipolar Recoupling (RFDR) spectra for the
ligand are presented. Hyperfine signals were well spread and
did not suffer from any sizable line broadening. The produc-
tion of H
1
receptor for Magic Angle Spinning NMR studies
is currently in progress.
Treatment: An agonist binding domain is proposed using
homology modeling, database searches and mutagenesis data
for the H
1
receptor.
Methods: Homology modeling, Database searches for Ex-
pressed sequence Tag (ESTs), Magic Angle Spinning Nuclear
Magnetic Resonance analysis of the ligand histamine.
Results: The three-dimensional receptor model and mutage-
nesis studies suggest that the amine of the agonist histamine
may form an ion pair with the TM III Asp, whereas the imi-
dazole ring of histamine may associate with TM V Asp and
Thr.
Conclusions: Homology modeling studies confirms the
absence of TM VIII in the H
1
receptor. According to the mod-
el the histamine in particular interacts with the transmem-
brane (TM) regions of the H
1
receptor structure, in particular
TM helix III and V. This is in line with recent mutagenesis
studies. Database search methods for ESTs have been used
for electronic prediction of tissue distribution of H
1
receptor
expression. The results indicate that the H
1
expression is
highest in heart and skeletal muscle, which may be of impor-
tance for drug targeting.
Key words: Histamine H
1
receptors – G protein coupled
receptors – MAS NMR – Ligand binding – ESTs
Inflamm. res. 52 (2003) 417 – 423
1023-3830/03/100417-07
DOI 10.1007/s00011-003-1195-3
Introduction
The development of selective histamine receptor(s) agonists
and antagonists has received considerable attention because
of their potential use as pharmacological agents against a
variety of human pathologies, including acid-peptic disor-
ders, duodenal ulcers, asthma and allergies. Biotechnology
using molecular biology, biophysics, and computational
approaches provides an alternative approach for classical
pharmacological screening to look at ligand-receptor interac-
tions and receptor specificity, which should allow to design
selective drugs based on detailed structural principles. Cur-
rent therapies as described in a standard pharmacology text,
for example Goodman & Gilman’s ‘The Pharmacological
Basis of Therapeutics’, are based on no more than 500 mole-
cular drug targets. These traditional targets reveal what have
been the most fruitful paths for therapeutic development in
the past, and give a glimpse of where genomic sciences may
yield drug-discovery success in the future. The cellular tar-
gets of most prescription drugs are proteins and a large pro-
portion of these are plasma membrane bound. For the latter
class, a drug does not need to enter the cell, but binds to the
extracellular binding site of the protein and can control intra-
cellular reactions from the outside. These molecular drug tar-
gets can be further categorized into biochemical classes such
as receptors, enzymes, matrix adhesion factors and ion chan-
nels. In this report we will address specific approaches to
study function, structure and relevance of a major pharma-
ceutical target family, namely G-Protein Coupled Receptors,
exemplified in the histamine H
1
receptor.
G-Protein Coupled Receptors
Biomembranes are based upon a bilayer matrix composed of
a mixture of lipid and other amphiphilic molecules creating
a thin but crucial separation between the inside and the out-
© Birkhäuser Verlag, Basel, 2003
Inflammation Research
Analysis of histamine and modeling of ligand-receptor
interactions in the histamine H
1
receptor for Magic Angle
Spinning NMR studies
V. R. Prasad Ratnala
1
, F. B. Hulsbergen
2
, H. J. M. de Groot
1
and W. J. de Grip
1,2
1
Leiden Institute of Chemistry, Gorlaeus Laboratoria, Einsteinweg 55, P.O. Box 9502, 2300 RA Leiden, The Netherlands, Fax: ++ 31 71 5274603,
e-mail: r.prasad@fwncism1.leidenuniv.nl
2
Department of Biochemistry UMC-160, Nijmegen Center of Molecular Life Sciences, University Medical Center Nijmegen, P.O. Box 9101,
6500 HB Nijmegen, The Netherlands
Received 3 June 2003; accepted by A. Falus 3 June 2003
Correspondence to: V. R. Prasad Ratnala