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Identification of aptamer-binding sites in hepatitis C virus envelope glycoprotein e2.

Chen F, Chen SC, Zhou J, Chen ZD, Chen F - Iran J Med Sci (2015)

Bottom Line: The peptide with high affinity was then further truncated, detected and compared with six kinds of HCV genotypes.The basic amino acid in 500 aa bound to ZE2 with high affinity, while acidic amino acid in 501 aa reduced the reaction between E2 and ZE2.The results showed the 500 aa and 501 aa of E2 were the key sites that bound to ZE2.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Life Sciences School of Hubei University, Wuhan, China;

ABSTRACT
Hepatitis C Virus (HCV) encodes two envelope glycoproteins, E1 and E2. Our previous work selected a specific aptamer ZE2, which could bind to E2 with high affinity, with a great potential for developing new molecular probes as an early diagnostic reagents or therapeutic drugs targeting HCV. In this study, the binding sites between E2 and aptamer ZE2 were further explored. E2 was truncated to 15 peptides (P1 to P15) and these peptides were used to detect the affinity with ZE2 by ELISA respectively. The peptide with high affinity was then further truncated, detected and compared with six kinds of HCV genotypes. The basic amino acid in 500 aa bound to ZE2 with high affinity, while acidic amino acid in 501 aa reduced the reaction between E2 and ZE2. The results showed the 500 aa and 501 aa of E2 were the key sites that bound to ZE2.

No MeSH data available.


Related in: MedlinePlus

Fluorescence microscope imaging of E2-expressing cells with FITC-ZE2 and FITC-ZE2-mut. A) FITC-ZE2 bound to the surface of E2-CT26 cells by confocal immunofluorescence microscopy. B) FITC-ZE2-mut was intracellular of the E2-CT26 cells by confocal immunofluorescence microscopy. Each test has repeated six times.
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Figure 1: Fluorescence microscope imaging of E2-expressing cells with FITC-ZE2 and FITC-ZE2-mut. A) FITC-ZE2 bound to the surface of E2-CT26 cells by confocal immunofluorescence microscopy. B) FITC-ZE2-mut was intracellular of the E2-CT26 cells by confocal immunofluorescence microscopy. Each test has repeated six times.

Mentions: The E2-expressing cells, E2-CT26, specifically bound FITC-ZE2 aptamer under the fluorescence microscope, indicating the affinity between synthesized aptamers and E2. The fluorescence intensities of the binding affinities between FITC-ZE2 and E2-expressing cells were higher than that between FITC-ZE2-mut and E2-expressing cells. Interestingly, we further observed ZE2-mut could enter into the cells, while ZE2 could only bind to the cells on the surface (figure 1). Previous studies reported that the synthesized aptamer ZE2 had similar function10 and the difference of ZE2 and ZE2-mut binding to E2-expressing cells was closely related to their structures.


Identification of aptamer-binding sites in hepatitis C virus envelope glycoprotein e2.

Chen F, Chen SC, Zhou J, Chen ZD, Chen F - Iran J Med Sci (2015)

Fluorescence microscope imaging of E2-expressing cells with FITC-ZE2 and FITC-ZE2-mut. A) FITC-ZE2 bound to the surface of E2-CT26 cells by confocal immunofluorescence microscopy. B) FITC-ZE2-mut was intracellular of the E2-CT26 cells by confocal immunofluorescence microscopy. Each test has repeated six times.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4300483&req=5

Figure 1: Fluorescence microscope imaging of E2-expressing cells with FITC-ZE2 and FITC-ZE2-mut. A) FITC-ZE2 bound to the surface of E2-CT26 cells by confocal immunofluorescence microscopy. B) FITC-ZE2-mut was intracellular of the E2-CT26 cells by confocal immunofluorescence microscopy. Each test has repeated six times.
Mentions: The E2-expressing cells, E2-CT26, specifically bound FITC-ZE2 aptamer under the fluorescence microscope, indicating the affinity between synthesized aptamers and E2. The fluorescence intensities of the binding affinities between FITC-ZE2 and E2-expressing cells were higher than that between FITC-ZE2-mut and E2-expressing cells. Interestingly, we further observed ZE2-mut could enter into the cells, while ZE2 could only bind to the cells on the surface (figure 1). Previous studies reported that the synthesized aptamer ZE2 had similar function10 and the difference of ZE2 and ZE2-mut binding to E2-expressing cells was closely related to their structures.

Bottom Line: The peptide with high affinity was then further truncated, detected and compared with six kinds of HCV genotypes.The basic amino acid in 500 aa bound to ZE2 with high affinity, while acidic amino acid in 501 aa reduced the reaction between E2 and ZE2.The results showed the 500 aa and 501 aa of E2 were the key sites that bound to ZE2.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Life Sciences School of Hubei University, Wuhan, China;

ABSTRACT
Hepatitis C Virus (HCV) encodes two envelope glycoproteins, E1 and E2. Our previous work selected a specific aptamer ZE2, which could bind to E2 with high affinity, with a great potential for developing new molecular probes as an early diagnostic reagents or therapeutic drugs targeting HCV. In this study, the binding sites between E2 and aptamer ZE2 were further explored. E2 was truncated to 15 peptides (P1 to P15) and these peptides were used to detect the affinity with ZE2 by ELISA respectively. The peptide with high affinity was then further truncated, detected and compared with six kinds of HCV genotypes. The basic amino acid in 500 aa bound to ZE2 with high affinity, while acidic amino acid in 501 aa reduced the reaction between E2 and ZE2. The results showed the 500 aa and 501 aa of E2 were the key sites that bound to ZE2.

No MeSH data available.


Related in: MedlinePlus