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In silico pharmacology2017; 5(1); 5; doi: 10.1007/s40203-017-0025-1

Molecular characterization, modeling, in silico analysis of equine pituitary gonadotropin alpha subunit and docking interaction studies with ganirelix.

Abstract: Equine pituitary gonadotropins (eLH, eFSH, eCG) are heterodimeric glycoprotein hormones with alpha (α) and beta (β) subunits. It is responsible for maintenance of pregnancy in mares during early gestation and fairly valuable for inducing superovulation in animals other than equines. The alpha subunit is common, while beta subunit is species-specific in all glycoprotein hormones. In the present investigation, molecular cloning and in silico characterization including homology modeling and molecular docking analysis of the equine chorionic gonadotropin (eCG) alpha subunit was carried out for gaining structural and functional insights into the eCG alpha subunit and its possible interaction with ganirelix, a gonadotropin-releasing hormone (GnRH) antagonist. The equine chorionic gonadotropin (eCG) alpha subunit expressed in pituitary gland was selected, amplified from total RNA, cloned and sequenced. The in silico analyses were made for homology modelling, structural details, epitope identification and chromosomal localization. Molecular docking studies of eCG alpha were undertaken with a drug ganirelix which is used to control ovulation and has antagonistic activity against GnRH. The protein sequence corresponding to selected open reading frame (ORF) was 99-100% similar with domesticated horse, Przewalski's horse, and 92-93% with Burchell's zebra and donkey. Molecular docking studies revealed the possible interaction of eCG alpha with ganirelix. The possible drug-macromolecule interactions were visualized between eCG alpha and ganirelix. The study will provide structural insight into unique sites and an alternate route of gonadotropin suppression applicable to assisted reproductive technologies.
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Publication Date: 2017-07-18 PubMed ID: 28721542PubMed Central: PMC5515723DOI: 10.1007/s40203-017-0025-1Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research article explores the molecular structure and function of the alpha subunit of equine pituitary gonadotropins, and its potential interaction with the hormone antagonist, ganirelix. The study offers significant insights into the possibilities of new methods in reproductive assistance.

Molecular Cloning

  • The alpha subunit of equine pituitary gonadotropins (eLH, eFSH, eCG) which are responsible for maintaining early-stage pregnancy in mares, was selected for this study. The choice is because it is common in all glycoprotein hormones, while the beta subunit is species-specific.
  • This alpha subunit was copied or amplified from the total RNA using the process of molecular cloning. Following this, it was sequenced.
  • The protein sequence turned out to resemble the sequences in domesticated horse, Przewalski’s horse, with a slight difference in Burchell’s zebra and donkey.

In Silico Analysis and Homology Modeling

  • Advanced computer-aided analysis was carried out for homology modelling – a way to create an atomic-resolution model of the “target” protein from its amino acid sequence and an experimental three-dimensional structure of a related homologous protein.
  • This analysis provided the researchers with detailed structural information, enabled antigen site identification, and identified chromosomal localization, articulating the position of the gene on the chromosome.

Docking Interaction with Ganirelix

  • A significant part of this study was molecular docking, which refers to the process of finding the compatible binding modes of a small molecule, such as a drug within the larger protein. In this case, ganirelix – a popular ovarian stimulant hormone antagonist, was chosen.
  • Analysis of docking interaction here revealed how the alpha subunit of eCG possibly interacts with ganirelix.
  • Peering into this interaction provided the researchers with the possibility of visualizing drug-macromolecule interactions between the alpha subunit of eCG and ganirelix.

Implications of the Study

  • The outcomes of the study provide significant structural insight into unique sites and an alternate route of gonadotropin suppression.
  • This could be of high importance to assisted reproductive technologies, providing a new way to control ovulation and manage reproduction.

Cite This Article

APA
Bhardwaj A, Nayan V, Sharma P, Kumar S, Pal Y, Singh J. (2017). Molecular characterization, modeling, in silico analysis of equine pituitary gonadotropin alpha subunit and docking interaction studies with ganirelix. In Silico Pharmacol, 5(1), 5. https://doi.org/10.1007/s40203-017-0025-1

Publication

In silico pharmacology
ISSN: 2193-9616
NlmUniqueID: 101623954
Country: Germany
Language: English
Volume: 5
Issue: 1
Pages: 5

Researcher Affiliations

Bhardwaj, Anuradha
  • Basic and Supporting Discipline Unit (BSDU), ICAR-National Research Centre on Equines, Hisar, Haryana, 125001, India. dranu.biotech@gmail.com.
Nayan, Varij
  • Animal Physiology and Reproduction (APR) Division, ICAR-Central Institute for Research on Buffaloes, Hisar, Haryana, 125001, India.
Sharma, Parvati
  • Basic and Supporting Discipline Unit (BSDU), ICAR-National Research Centre on Equines, Hisar, Haryana, 125001, India.
Kumar, Sanjay
  • Equine Health Unit (EHU), ICAR-National Research Centre on Equines, Hisar, Haryana, 125001, India.
Pal, Yash
  • Equine Production Centre, ICAR-National Research Centre on Equines, Bikaner, Rajasthan, 334 001, India.
Singh, Jitender
  • Equine Production Centre, ICAR-National Research Centre on Equines, Bikaner, Rajasthan, 334 001, India.

Conflict of Interest Statement

STATEMENT OF ETHICS APPROVAL: Experimental protocol was reviewed and approved by Institute Animal Ethics Committee (IAEC) of ICAR-National Research Centre on Equines, Hisar. All the procedures for sample collection were performed according to the approval by the Institute Animal Ethics Committee (IAEC) of ICAR-National Research Centre on Equines vide NRCE/CPCSEA/2012-13/dated the 18th July 2012. CONSENT TO PARTICIPATE: We give our consent to participate under the ‘Ethics, consent and permissions’ heading. AVAILABILITY OF DATA AND MATERIALS: The data generated during this study and supporting this study are included in this article and also in additional files within the Additional file 1: Figure S1–S11 and Additional file 2: tables S1-S2. The dataset analyzed during the current study is available from the corresponding author on a reasonable request. CONFLICT OF INTEREST: None of the authors have any competing interests in the manuscript. FUNDING: This work was financially supported by ICAR-National Research Centre on Equines, Hisar (URL: www.nrce.gov.in) under the institute project (code: IXX02769).

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