Spectroscopic evaluation of synthesized 5β-dihydrocortisol and 5β-dihydrocortisol acetate binding mechanism with human serum albumin and their role in anticancer activity

Monika Kallubai, Srinivasa P. Reddy, Shreya Dubey, Dhevalapally B. Ramachary, S. Rajagopal

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Our study focus on the biological importance of synthesized 5β-dihydrocortisol (Dhc) and 5β-dihydrocortisol acetate (DhcA) molecules, the cytotoxic study was performed on breast cancer cell line (MCF-7) normal human embryonic kidney cell line (HEK293), the IC50 values for MCF-7 cells were 28 and 25 μM, respectively, whereas no toxicity in terms of cell viability was observed with HEK293 cell line. Further experiment proved that Dhc and DhcA induced 35.6 and 37.7% early apoptotic cells and 2.5, 2.9% late apoptotic cells, respectively, morphological observation of cell death through TUNEL assay revealed that Dhc and DhcA induced apoptosis in MCF-7 cells. The complexes of HSA–Dhc and HSA–DhcA were observed as static quenching, and the binding constants (K) was 4.7 ± .03 × 104 M−1 and 3.9 ± .05 × 104 M−1 , and their binding free energies were found to be −6.4 and −6.16 kcal/mol, respectively. The displacement studies confirmed that lidocaine 1.4 ± .05 × 104 M−1 replaced Dhc, and phenylbutazone 1.5 ± .05 × 104 M−1 replaced by DhcA, which explains domain I and domain II are the binding sites for Dhc and DhcA. Further, FT-IR, synchronous spectroscopy, and CD results revealed that the secondary structure of HSA was altered in the presence of Dhc and DhcA. Furthermore, the atomic force microscopy and transmission electron microscopy showed that the dimensions like height and molecular size of the HSA–Dhc and HSA–DhcA complex were larger compared to HSA alone. Detailed analysis through molecular dynamics simulations also supported greater stability of HSA–Dhc and HSA–DhcA complexes, and root-mean-square-fluctuation interpreted the binding site of Dhc as domain IB and domain IIA for DhcA. This information is valuable for further development of steroid derivative with improved pharmacological significance as novel anti-cancer drugs.

Original languageEnglish
Pages (from-to)1-18
Number of pages18
JournalJournal of Biomolecular Structure and Dynamics
DOIs
Publication statusAccepted/In press - Feb 15 2018

Fingerprint

Serum Albumin
Acetates
MCF-7 Cells
5-dihydrocortisol
Cell Line
Binding Sites
Phenylbutazone
Atomic Force Microscopy
HEK293 Cells
In Situ Nick-End Labeling
Molecular Dynamics Simulation
Lidocaine
Transmission Electron Microscopy
Inhibitory Concentration 50
Spectrum Analysis
Cell Survival
Cell Death
Steroids
Observation
Pharmacology

Keywords

  • 5β-dihydrocortisol
  • apoptosis
  • fluorescence quenching
  • molecular docking
  • molecular dynamics simulations
  • protein conformations

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Cite this

Spectroscopic evaluation of synthesized 5β-dihydrocortisol and 5β-dihydrocortisol acetate binding mechanism with human serum albumin and their role in anticancer activity. / Kallubai, Monika; Reddy, Srinivasa P.; Dubey, Shreya; Ramachary, Dhevalapally B.; Rajagopal, S.

In: Journal of Biomolecular Structure and Dynamics, 15.02.2018, p. 1-18.

Research output: Contribution to journalArticle

@article{901e8e5c0bff4188b713228271595138,
title = "Spectroscopic evaluation of synthesized 5β-dihydrocortisol and 5β-dihydrocortisol acetate binding mechanism with human serum albumin and their role in anticancer activity",
abstract = "Our study focus on the biological importance of synthesized 5β-dihydrocortisol (Dhc) and 5β-dihydrocortisol acetate (DhcA) molecules, the cytotoxic study was performed on breast cancer cell line (MCF-7) normal human embryonic kidney cell line (HEK293), the IC50 values for MCF-7 cells were 28 and 25 μM, respectively, whereas no toxicity in terms of cell viability was observed with HEK293 cell line. Further experiment proved that Dhc and DhcA induced 35.6 and 37.7{\%} early apoptotic cells and 2.5, 2.9{\%} late apoptotic cells, respectively, morphological observation of cell death through TUNEL assay revealed that Dhc and DhcA induced apoptosis in MCF-7 cells. The complexes of HSA–Dhc and HSA–DhcA were observed as static quenching, and the binding constants (K) was 4.7 ± .03 × 104 M−1 and 3.9 ± .05 × 104 M−1 , and their binding free energies were found to be −6.4 and −6.16 kcal/mol, respectively. The displacement studies confirmed that lidocaine 1.4 ± .05 × 104 M−1 replaced Dhc, and phenylbutazone 1.5 ± .05 × 104 M−1 replaced by DhcA, which explains domain I and domain II are the binding sites for Dhc and DhcA. Further, FT-IR, synchronous spectroscopy, and CD results revealed that the secondary structure of HSA was altered in the presence of Dhc and DhcA. Furthermore, the atomic force microscopy and transmission electron microscopy showed that the dimensions like height and molecular size of the HSA–Dhc and HSA–DhcA complex were larger compared to HSA alone. Detailed analysis through molecular dynamics simulations also supported greater stability of HSA–Dhc and HSA–DhcA complexes, and root-mean-square-fluctuation interpreted the binding site of Dhc as domain IB and domain IIA for DhcA. This information is valuable for further development of steroid derivative with improved pharmacological significance as novel anti-cancer drugs.",
keywords = "5β-dihydrocortisol, apoptosis, fluorescence quenching, molecular docking, molecular dynamics simulations, protein conformations",
author = "Monika Kallubai and Reddy, {Srinivasa P.} and Shreya Dubey and Ramachary, {Dhevalapally B.} and S. Rajagopal",
year = "2018",
month = "2",
day = "15",
doi = "10.1080/07391102.2018.1433554",
language = "English",
pages = "1--18",
journal = "Journal of Biomolecular Structure and Dynamics",
issn = "0739-1102",
publisher = "Adenine Press",

}

TY - JOUR

T1 - Spectroscopic evaluation of synthesized 5β-dihydrocortisol and 5β-dihydrocortisol acetate binding mechanism with human serum albumin and their role in anticancer activity

AU - Kallubai, Monika

AU - Reddy, Srinivasa P.

AU - Dubey, Shreya

AU - Ramachary, Dhevalapally B.

AU - Rajagopal, S.

PY - 2018/2/15

Y1 - 2018/2/15

N2 - Our study focus on the biological importance of synthesized 5β-dihydrocortisol (Dhc) and 5β-dihydrocortisol acetate (DhcA) molecules, the cytotoxic study was performed on breast cancer cell line (MCF-7) normal human embryonic kidney cell line (HEK293), the IC50 values for MCF-7 cells were 28 and 25 μM, respectively, whereas no toxicity in terms of cell viability was observed with HEK293 cell line. Further experiment proved that Dhc and DhcA induced 35.6 and 37.7% early apoptotic cells and 2.5, 2.9% late apoptotic cells, respectively, morphological observation of cell death through TUNEL assay revealed that Dhc and DhcA induced apoptosis in MCF-7 cells. The complexes of HSA–Dhc and HSA–DhcA were observed as static quenching, and the binding constants (K) was 4.7 ± .03 × 104 M−1 and 3.9 ± .05 × 104 M−1 , and their binding free energies were found to be −6.4 and −6.16 kcal/mol, respectively. The displacement studies confirmed that lidocaine 1.4 ± .05 × 104 M−1 replaced Dhc, and phenylbutazone 1.5 ± .05 × 104 M−1 replaced by DhcA, which explains domain I and domain II are the binding sites for Dhc and DhcA. Further, FT-IR, synchronous spectroscopy, and CD results revealed that the secondary structure of HSA was altered in the presence of Dhc and DhcA. Furthermore, the atomic force microscopy and transmission electron microscopy showed that the dimensions like height and molecular size of the HSA–Dhc and HSA–DhcA complex were larger compared to HSA alone. Detailed analysis through molecular dynamics simulations also supported greater stability of HSA–Dhc and HSA–DhcA complexes, and root-mean-square-fluctuation interpreted the binding site of Dhc as domain IB and domain IIA for DhcA. This information is valuable for further development of steroid derivative with improved pharmacological significance as novel anti-cancer drugs.

AB - Our study focus on the biological importance of synthesized 5β-dihydrocortisol (Dhc) and 5β-dihydrocortisol acetate (DhcA) molecules, the cytotoxic study was performed on breast cancer cell line (MCF-7) normal human embryonic kidney cell line (HEK293), the IC50 values for MCF-7 cells were 28 and 25 μM, respectively, whereas no toxicity in terms of cell viability was observed with HEK293 cell line. Further experiment proved that Dhc and DhcA induced 35.6 and 37.7% early apoptotic cells and 2.5, 2.9% late apoptotic cells, respectively, morphological observation of cell death through TUNEL assay revealed that Dhc and DhcA induced apoptosis in MCF-7 cells. The complexes of HSA–Dhc and HSA–DhcA were observed as static quenching, and the binding constants (K) was 4.7 ± .03 × 104 M−1 and 3.9 ± .05 × 104 M−1 , and their binding free energies were found to be −6.4 and −6.16 kcal/mol, respectively. The displacement studies confirmed that lidocaine 1.4 ± .05 × 104 M−1 replaced Dhc, and phenylbutazone 1.5 ± .05 × 104 M−1 replaced by DhcA, which explains domain I and domain II are the binding sites for Dhc and DhcA. Further, FT-IR, synchronous spectroscopy, and CD results revealed that the secondary structure of HSA was altered in the presence of Dhc and DhcA. Furthermore, the atomic force microscopy and transmission electron microscopy showed that the dimensions like height and molecular size of the HSA–Dhc and HSA–DhcA complex were larger compared to HSA alone. Detailed analysis through molecular dynamics simulations also supported greater stability of HSA–Dhc and HSA–DhcA complexes, and root-mean-square-fluctuation interpreted the binding site of Dhc as domain IB and domain IIA for DhcA. This information is valuable for further development of steroid derivative with improved pharmacological significance as novel anti-cancer drugs.

KW - 5β-dihydrocortisol

KW - apoptosis

KW - fluorescence quenching

KW - molecular docking

KW - molecular dynamics simulations

KW - protein conformations

UR - http://www.scopus.com/inward/record.url?scp=85042066108&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85042066108&partnerID=8YFLogxK

U2 - 10.1080/07391102.2018.1433554

DO - 10.1080/07391102.2018.1433554

M3 - Article

SP - 1

EP - 18

JO - Journal of Biomolecular Structure and Dynamics

JF - Journal of Biomolecular Structure and Dynamics

SN - 0739-1102

ER -