Structurally Different Yet Functionally Similar: Aptamers Specific for the Ebola Virus Soluble Glycoprotein and GP1,2 and Their Application in Electrochemical Sensing
The Ebola virus glycoprotein (GP) gene templates several mRNAs that produce either the virion-associated transmembrane protein or one of two secreted glycoproteins. Soluble glycoprotein (sGP) is the predominant product. GP1 and sGP share an amino terminal sequence of 295 amino acids but differ in qu...
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-02-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/24/5/4627 |
| _version_ | 1797615196042166272 |
|---|---|
| author | Soma Banerjee Mahsa Askary Hemmat Shambhavi Shubham Agnivo Gosai Sivaranjani Devarakonda Nianyu Jiang Charith Geekiyanage Jacob A. Dillard Wendy Maury Pranav Shrotriya Monica H. Lamm Marit Nilsen-Hamilton |
| author_facet | Soma Banerjee Mahsa Askary Hemmat Shambhavi Shubham Agnivo Gosai Sivaranjani Devarakonda Nianyu Jiang Charith Geekiyanage Jacob A. Dillard Wendy Maury Pranav Shrotriya Monica H. Lamm Marit Nilsen-Hamilton |
| author_sort | Soma Banerjee |
| collection | DOAJ |
| description | The Ebola virus glycoprotein (GP) gene templates several mRNAs that produce either the virion-associated transmembrane protein or one of two secreted glycoproteins. Soluble glycoprotein (sGP) is the predominant product. GP1 and sGP share an amino terminal sequence of 295 amino acids but differ in quaternary structure, with GP1 being a heterohexamer with GP2 and sGP a homodimer. Two structurally different DNA aptamers were selected against sGP that also bound GP1,2. These DNA aptamers were compared with a 2′FY-RNA aptamer for their interactions with the Ebola GP gene products. The three aptamers have almost identical binding isotherms for sGP and GP1,2 in solution and on the virion. They demonstrated high affinity and selectivity for sGP and GP1,2. Furthermore, one aptamer, used as a sensing element in an electrochemical format, detected GP1,2 on pseudotyped virions and sGP with high sensitivity in the presence of serum, including from an Ebola-virus-infected monkey. Our results suggest that the aptamers interact with sGP across the interface between the monomers, which is different from the sites on the protein bound by most antibodies. The remarkable similarity in functional features of three structurally distinct aptamers suggests that aptamers, like antibodies, have preferred binding sites on proteins. |
| first_indexed | 2024-03-11T07:22:57Z |
| format | Article |
| id | doaj.art-c60b75daa65f416b92021e7b4ca26610 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T07:22:57Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-c60b75daa65f416b92021e7b4ca266102023-11-17T07:51:10ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-02-01245462710.3390/ijms24054627Structurally Different Yet Functionally Similar: Aptamers Specific for the Ebola Virus Soluble Glycoprotein and GP1,2 and Their Application in Electrochemical SensingSoma Banerjee0Mahsa Askary Hemmat1Shambhavi Shubham2Agnivo Gosai3Sivaranjani Devarakonda4Nianyu Jiang5Charith Geekiyanage6Jacob A. Dillard7Wendy Maury8Pranav Shrotriya9Monica H. Lamm10Marit Nilsen-Hamilton11Ames Laboratory, U.S. Department of Energy, Ames, IA 50011, USARoy J. Carver Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, IA 50011, USARoy J. Carver Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, IA 50011, USADepartment of Mechanical Engineering, Iowa State University, Ames, IA 50011, USADepartment of Mechanical Engineering, Iowa State University, Ames, IA 50011, USADepartment of Mechanical Engineering, Iowa State University, Ames, IA 50011, USAAptalogic Inc., Ames, IA 50014, USADepartment of Microbiology and Immunology, University of Iowa, Iowa City, IA 50011, USADepartment of Microbiology and Immunology, University of Iowa, Iowa City, IA 50011, USADepartment of Mechanical Engineering, Iowa State University, Ames, IA 50011, USADepartment of Chemical and Biological Engineering, Iowa State University, Ames, IA 50011, USAAmes Laboratory, U.S. Department of Energy, Ames, IA 50011, USAThe Ebola virus glycoprotein (GP) gene templates several mRNAs that produce either the virion-associated transmembrane protein or one of two secreted glycoproteins. Soluble glycoprotein (sGP) is the predominant product. GP1 and sGP share an amino terminal sequence of 295 amino acids but differ in quaternary structure, with GP1 being a heterohexamer with GP2 and sGP a homodimer. Two structurally different DNA aptamers were selected against sGP that also bound GP1,2. These DNA aptamers were compared with a 2′FY-RNA aptamer for their interactions with the Ebola GP gene products. The three aptamers have almost identical binding isotherms for sGP and GP1,2 in solution and on the virion. They demonstrated high affinity and selectivity for sGP and GP1,2. Furthermore, one aptamer, used as a sensing element in an electrochemical format, detected GP1,2 on pseudotyped virions and sGP with high sensitivity in the presence of serum, including from an Ebola-virus-infected monkey. Our results suggest that the aptamers interact with sGP across the interface between the monomers, which is different from the sites on the protein bound by most antibodies. The remarkable similarity in functional features of three structurally distinct aptamers suggests that aptamers, like antibodies, have preferred binding sites on proteins.https://www.mdpi.com/1422-0067/24/5/4627aptamersEbola GP1,2Ebola sGPelectrochemical sensoraptamer–protein interaction |
| spellingShingle | Soma Banerjee Mahsa Askary Hemmat Shambhavi Shubham Agnivo Gosai Sivaranjani Devarakonda Nianyu Jiang Charith Geekiyanage Jacob A. Dillard Wendy Maury Pranav Shrotriya Monica H. Lamm Marit Nilsen-Hamilton Structurally Different Yet Functionally Similar: Aptamers Specific for the Ebola Virus Soluble Glycoprotein and GP1,2 and Their Application in Electrochemical Sensing International Journal of Molecular Sciences aptamers Ebola GP1,2 Ebola sGP electrochemical sensor aptamer–protein interaction |
| title | Structurally Different Yet Functionally Similar: Aptamers Specific for the Ebola Virus Soluble Glycoprotein and GP1,2 and Their Application in Electrochemical Sensing |
| title_full | Structurally Different Yet Functionally Similar: Aptamers Specific for the Ebola Virus Soluble Glycoprotein and GP1,2 and Their Application in Electrochemical Sensing |
| title_fullStr | Structurally Different Yet Functionally Similar: Aptamers Specific for the Ebola Virus Soluble Glycoprotein and GP1,2 and Their Application in Electrochemical Sensing |
| title_full_unstemmed | Structurally Different Yet Functionally Similar: Aptamers Specific for the Ebola Virus Soluble Glycoprotein and GP1,2 and Their Application in Electrochemical Sensing |
| title_short | Structurally Different Yet Functionally Similar: Aptamers Specific for the Ebola Virus Soluble Glycoprotein and GP1,2 and Their Application in Electrochemical Sensing |
| title_sort | structurally different yet functionally similar aptamers specific for the ebola virus soluble glycoprotein and gp1 2 and their application in electrochemical sensing |
| topic | aptamers Ebola GP1,2 Ebola sGP electrochemical sensor aptamer–protein interaction |
| url | https://www.mdpi.com/1422-0067/24/5/4627 |
| work_keys_str_mv | AT somabanerjee structurallydifferentyetfunctionallysimilaraptamersspecificfortheebolavirussolubleglycoproteinandgp12andtheirapplicationinelectrochemicalsensing AT mahsaaskaryhemmat structurallydifferentyetfunctionallysimilaraptamersspecificfortheebolavirussolubleglycoproteinandgp12andtheirapplicationinelectrochemicalsensing AT shambhavishubham structurallydifferentyetfunctionallysimilaraptamersspecificfortheebolavirussolubleglycoproteinandgp12andtheirapplicationinelectrochemicalsensing AT agnivogosai structurallydifferentyetfunctionallysimilaraptamersspecificfortheebolavirussolubleglycoproteinandgp12andtheirapplicationinelectrochemicalsensing AT sivaranjanidevarakonda structurallydifferentyetfunctionallysimilaraptamersspecificfortheebolavirussolubleglycoproteinandgp12andtheirapplicationinelectrochemicalsensing AT nianyujiang structurallydifferentyetfunctionallysimilaraptamersspecificfortheebolavirussolubleglycoproteinandgp12andtheirapplicationinelectrochemicalsensing AT charithgeekiyanage structurallydifferentyetfunctionallysimilaraptamersspecificfortheebolavirussolubleglycoproteinandgp12andtheirapplicationinelectrochemicalsensing AT jacobadillard structurallydifferentyetfunctionallysimilaraptamersspecificfortheebolavirussolubleglycoproteinandgp12andtheirapplicationinelectrochemicalsensing AT wendymaury structurallydifferentyetfunctionallysimilaraptamersspecificfortheebolavirussolubleglycoproteinandgp12andtheirapplicationinelectrochemicalsensing AT pranavshrotriya structurallydifferentyetfunctionallysimilaraptamersspecificfortheebolavirussolubleglycoproteinandgp12andtheirapplicationinelectrochemicalsensing AT monicahlamm structurallydifferentyetfunctionallysimilaraptamersspecificfortheebolavirussolubleglycoproteinandgp12andtheirapplicationinelectrochemicalsensing AT maritnilsenhamilton structurallydifferentyetfunctionallysimilaraptamersspecificfortheebolavirussolubleglycoproteinandgp12andtheirapplicationinelectrochemicalsensing |