Vol 11, Special issue 2, 2018
Online - 2455-3891
Print - 0974-2441
STRUCTURAL CHARACTERISATION OF 5-HYDROXYTRYPTAMINE
2A
RECEPTOR IN HOMO
SAPIENS BY IN - SILICO METHOD
SONI SINGH, ALOK JHA*
Department of Biotechnology and Life Sciences, Mangalayatan University, Aligarh, Uttar Pradesh, India. Email: alok.gene@gmail.com
Received: 19 March 2018, Revised and Accepted: 10 July 2018
ABSTRACT
Objective: Structural characterization of 5-hydroxytryptamine (5-HT)
2A
receptor in homo sapiens using in silico method.
Methods: In silico approach has particularly providing a realistic representation needed to understand the fundamental molecular structure of a
serotonin receptor. The structure has been generated using Swiss model, Modeller 9.14, Phyre2, and Geno three-dimensional, which was visualized
using PyMol, and validated by Procheck and ERRAT analysis along with the values of different secondary structures mapping to diverse sections of
the Ramachandran plot.
Results: We compared all different models. Further structural analysis suggested that the structure of 5-HT
2A
is a monomer with 18 alpha helices,
seven beta sheets, and one disulfide bridge. There is no signal peptide region in the protein sequence. The structure contains mostly polar and
aromatic amino acid as suggested by using hydropathy plot. However, in both partitioning systems bilayer to water and water to bilayer, there are
some hydropathy predicted segments, which are also transmembrane segments. Finally, the pore features, including diameter profile, size, and shape,
were determined by porewalker, and the shape of the pore was found to be UDSD.
Conclusion: This study suggested that 5-HT
2A
receptor interaction with its natural ligand serotonin and other inhibitor compounds would further
additional information about G protein-coupled receptors. The 5-HT
2A
receptor could be an important target for therapeutics development.
Keywords: 5-Hydroxytryptamine
2a
receptor, Homology modeling, G Protein-coupled receptors, Transmembrane protein, Model comparison.
INTRODUCTION
Serotonin 5-hydroxytryptamine (5-HT) is one of the neurotransmitters
present at synapses of nerve cells. In the central nervous system
(CNS), serotonin is mainly involved in the regulation of depression,
anxiety, aggression, memory, appetite, cognition, sleep, emotion,
perception, and consciousness [1]. The 5-HT receptors have been
classified into 5-HT
1
–5HT
7
, and again they are divided into 12 different
subpopulations. Some agonists and antagonists for subpopulations of
5-HT receptors are being developed by using different approaches.
Still as far yet, to design and develop a specific inhibitor for 5-HT
receptors with therapeutic potential is a challenge, although many
agonists and antagonists have been suggested [2]. All serotonergic
receptors come under the category of G-protein-coupled receptor
(GPCR) superfamily [3] except 5-HT3 receptor. The 5-HT3 receptor
is one of the ligand-gated ion channels and depends on the nicotinic
acetylcholine receptor superfamily having cysteine-loop transmitter
gate, and constitute of heteropentamers [4,5]. The 5-HT
2A
receptor
is mainly present in the prefrontal cortex of CNS, and they are
present in that region of the brain, which is essential for learning and
cognition. The activity of this receptor is linked to many neurological
disorders and conditions such as schizophrenia and depression. [5].
The 5HT
2
receptors are subdivided into 5-HT
2A
, 5-HT
2B
, and 5-HT
2C
receptors. 5-HT
2A
receptors exhibit high sequence homology with
other 5-HT receptors (78%). Phenylalkylamines (like (2,5-dimethyl-
4-bromoamphetamine) and (2,5-dimethoxy-4iodoamphetamine)) are
well known agonists for 5-HT
2A
[6]. There are so many drugs available
to target this receptor. Although the receptor has been widely studied
about multiple functions in the CNS, high level of this receptor has
been studied in other parts of the body such as platelets, endothelial
cells, and intestine (Fig. 1).
METHODS
Template selection and model building
The protein sequence and information of 5-HT
2A
were collected from
the National Center of Biotechnology (NCBI), in FASTA format, with the
accession number NP > AAH96839.1 5. The sequence length reported
to be 471 amino acids. To search for homologous sequence of 5-HT
2A
,
BLASTP was performed against the nonredundant database of NCBI.
Sequences are selected from the sequence similarity search based
on the identity percentage (cutoff 95% identity). The 471 amino acid
residue of 5-HT
2A
receptor was subjected to BLASTP against the Protein
Data Bank [7] to identify suitable template for comparative study
of the protein structure for modeling.Further, the protein sequence
was subjected to comparative homology modeling through Modeler
9.14, SWISS-MODEL Server [8], PHYRE2 server [9], and Geno three-
dimensional (3D) server [10]. These modeling servers on the basis of
their different algorithm provide 3D structure of 5-HT
2A
structure for
further study.
Validation of generated model
Finally, once the 3D structures were generated, evaluation of the
structure and stereochemical analysis was performed using different
types of validation tools. A study of backbone conformation of all
models was evaluated by the analysis of Ramachandran plot using
RAMPAGE program [11]. ERRAT tool finds the overall quality factor of
the proteins. ERRAT plot gives statistics of the error in the structure
for each residue in the 3D protein structure model. This process was
repeated, until when most of the amino acid residues was shown below
95% cutoff [12].
© 2018 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.
org/licenses/by/4. 0/) DOI: http://dx.doi.org/10.22159/ajpcr.2018.v11s2.28588
Research Article
Recent Trends in Biomedical Sciences-2018 (RTBS-2018)