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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

Identification of the ZE2-binding site of HCV E2 envelope glycoprotein. A) The binding affinity between aptamers and E2 fragment (from P1 to P15) was detected by indirect ELISA. B) Diagram of the 15 E2 peptide fragments (from P1 to P15) that spanned different regions of E2, and the P7 fragment was truncated to P7-1, P7-2, and P7-3. C) The binding affinity was detected between aptamers and three peptides of P7. Each test has been repeated three times. (**P<0.01)
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Figure 2: Identification of the ZE2-binding site of HCV E2 envelope glycoprotein. A) The binding affinity between aptamers and E2 fragment (from P1 to P15) was detected by indirect ELISA. B) Diagram of the 15 E2 peptide fragments (from P1 to P15) that spanned different regions of E2, and the P7 fragment was truncated to P7-1, P7-2, and P7-3. C) The binding affinity was detected between aptamers and three peptides of P7. Each test has been repeated three times. (**P<0.01)

Mentions: To explore the ZE2 binding site on E2, fifteen peptide fragments were synthesized, which were based on the genotype 1a E2 amino acid sequence and secondary structure (table 1). Those fifteen fragments were used to coat ELISA plates and incubated with biotin-labeled ZE2. The binding affinity was detected by ELISA, as described earlier. Among those fifteen fragments, the results suggest that ZE2 bound most strongly to the E2-P7 fragment sequence, while ZE2-mut showed slightly lower binding affinities than ZE2 (figure 2). The binding affinity between ZE2 and P7 was 2- to 20-fold higher compared with those between ZE2 and the other 14 peptides (P=0.00). Since ZE2 bound most strongly to the E2-P7 fragment sequence, we further truncated E2-P7 to three smaller fragments: P7-1, P7-2, and P7-3 (figure 2). The indirect ELISAs were used to detect the affinity between the aptamer and peptides (P7-1, P7-2, and P7-3). The results indicated that there is no significant difference of binding to peptides between ZE2 and ZE2-mut.


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)

Identification of the ZE2-binding site of HCV E2 envelope glycoprotein. A) The binding affinity between aptamers and E2 fragment (from P1 to P15) was detected by indirect ELISA. B) Diagram of the 15 E2 peptide fragments (from P1 to P15) that spanned different regions of E2, and the P7 fragment was truncated to P7-1, P7-2, and P7-3. C) The binding affinity was detected between aptamers and three peptides of P7. Each test has been repeated three times. (**P<0.01)
© Copyright Policy
Related In: Results  -  Collection

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

Figure 2: Identification of the ZE2-binding site of HCV E2 envelope glycoprotein. A) The binding affinity between aptamers and E2 fragment (from P1 to P15) was detected by indirect ELISA. B) Diagram of the 15 E2 peptide fragments (from P1 to P15) that spanned different regions of E2, and the P7 fragment was truncated to P7-1, P7-2, and P7-3. C) The binding affinity was detected between aptamers and three peptides of P7. Each test has been repeated three times. (**P<0.01)
Mentions: To explore the ZE2 binding site on E2, fifteen peptide fragments were synthesized, which were based on the genotype 1a E2 amino acid sequence and secondary structure (table 1). Those fifteen fragments were used to coat ELISA plates and incubated with biotin-labeled ZE2. The binding affinity was detected by ELISA, as described earlier. Among those fifteen fragments, the results suggest that ZE2 bound most strongly to the E2-P7 fragment sequence, while ZE2-mut showed slightly lower binding affinities than ZE2 (figure 2). The binding affinity between ZE2 and P7 was 2- to 20-fold higher compared with those between ZE2 and the other 14 peptides (P=0.00). Since ZE2 bound most strongly to the E2-P7 fragment sequence, we further truncated E2-P7 to three smaller fragments: P7-1, P7-2, and P7-3 (figure 2). The indirect ELISAs were used to detect the affinity between the aptamer and peptides (P7-1, P7-2, and P7-3). The results indicated that there is no significant difference of binding to peptides between ZE2 and ZE2-mut.

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