Title:An Intracellular Tripeptide Arg-His-Trp of Serum Origin Detected in MCF-7 Cells is a Possible Agonist to β2 Adrenoceptor
Volume: 28
Issue: 10
Author(s): Hritik Chandore, Ajay Kumar Raj, Kiran Bharat Lokhande, Krishna Venkateswara Swamy, Jayanta Kumar Pal and Nilesh Kumar Sharma*
Affiliation:
- Cancer and Translational Research Lab, Dr. D.Y. Patil Biotechnology & Bioinformatics Institute, Dr. D.Y. Patil Vidyapeeth, Pune, Maharashtra 411033,India
Keywords:
Metabolites, serum, tripeptides, breast cancer cells, agonists, adrenergic receptor, molecular dynamic simulations.
Abstract:
Background: The need for agonists and antagonists of β2 adrenoceptor (β2AR) is warranted
in various human disease conditions, including cancer, cardiovascular and other metabolic
disorders. However, the sources of agonists of β2AR are diverse in nature. Interestingly, there is a
complete gap in the exploration of agonists of β2AR from serum that is a well-known component
of culture media that supports growth and proliferation of normal and cancer cells in vitro.
Methods: In this paper, we employed a novel vertical tube gel electrophoresis (VTGE)-assisted purification
of intracellular metabolites of MCF-7 cells grown in vitro in complete media with fetal
bovine serum (FBS). Intracellular metabolites of MCF-7 cells were then analyzed by LC-HRMS.
Identified intracellular tripeptides of FBS origin were evaluated for their molecular interactions
with various extracellular and intracellular receptors, including β2AR (PDB ID: 2RH1) by employing
molecular docking and molecular dynamics simulations (MDS). A known agonist of β2AR, isoproterenol
was used as a positive control in molecular docking and MDS analyses.
Results: We report here the identification of a few novel intracellular tripeptides, namely Arg-His-
Trp, (PubChem CID-145453842), Pro-Ile-Glu, (PubChem CID-145457492), Cys-Gln-Gln,
(PubChem CID-71471965), Glu-Glu-Lys, (PubChem CID-11441068) and Gly-Cys-Leu (PubChem
CID-145455600) of FBS origin in MCF-7 cells. Molecular docking and MDS analyses revealed
that among these molecules, the tripeptide Arg-His-Trp shows a favorable binding affinity with
β2AR (-9.8 Kcal/mol). The agonistic effect of Arg-His-Trp is significant and comparable with that
of a known agonist of β2AR, isoproterenol.
Conclusion: In conclusion, we identified a unique Arg-His-Trp tripeptide of FBS origin in MCF-7
cells by employing a novel approach. This unique tripeptide Arg-His-Trp is suggested to be a potential
agonist of β2AR and it may have applications in the context of various human diseases like
bronchial asthma and chronic obstructive pulmonary disease (COPD).
