A Structural Bioinformatics-Guided Study of Adenosine Triphosphate-Binding Cassette (ABC) Transporters and Their Substrates
Adenosine triphosphate-binding cassette (ABC) transporters form a ubiquitous superfamily of integral membrane proteins involved in the translocation of substrates across membranes. Human ABC transporters are closely linked to the pathogenesis of diseases such as cancer, metabolic diseases, and Alzhe...
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MDPI AG
2025-01-01
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| Series: | Membranes |
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| Online Access: | https://www.mdpi.com/2077-0375/15/1/20 |
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| author | Iqra Younus Robert C. Ford Stephen M. Prince |
| author_facet | Iqra Younus Robert C. Ford Stephen M. Prince |
| author_sort | Iqra Younus |
| collection | DOAJ |
| description | Adenosine triphosphate-binding cassette (ABC) transporters form a ubiquitous superfamily of integral membrane proteins involved in the translocation of substrates across membranes. Human ABC transporters are closely linked to the pathogenesis of diseases such as cancer, metabolic diseases, and Alzheimer’s disease. In this study, four ABC transporters were chosen based on (I) their importance in humans and (II) their score in a structural bioinformatics screen aimed at the prediction of crystallisation propensity. The top-scoring ABC transporters’ orthologs (<i>Mus musculus</i>—mouse ABCB5, <i>Ailuropoda melanoleuca</i>—giant panda ABCB6, <i>Myotis lucifugus</i>—little brown bat ABCG1 and <i>Mus musculus</i> ABCG4) were then expressed in <i>Saccharomyces cerevisiae</i> with a combined green fluorescent protein and polyhistidine tag, enabling visualisation and purification. After partial purification and in the presence of the detergent (n-dodecyl-β-D-maltoside), the kinetic parameters of the ATP hydrolysis reactions of the orthologs were determined, as well as the extent of stimulation of their activity when presented with putative substrates. We discuss the efficiency of such bioinformatics approaches and make suggestions for their improvement and wider application in membrane protein-structure determination. |
| format | Article |
| id | doaj-art-8753b62fa7384cc592e4200cdd103b55 |
| institution | Kabale University |
| issn | 2077-0375 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Membranes |
| spelling | doaj-art-8753b62fa7384cc592e4200cdd103b552025-01-24T13:41:02ZengMDPI AGMembranes2077-03752025-01-011512010.3390/membranes15010020A Structural Bioinformatics-Guided Study of Adenosine Triphosphate-Binding Cassette (ABC) Transporters and Their SubstratesIqra Younus0Robert C. Ford1Stephen M. Prince2Faculty of Biology, Medicine and Health, School of Biological Sciences, The University of Manchester, Manchester M13 9PT, UKFaculty of Biology, Medicine and Health, School of Biological Sciences, The University of Manchester, Manchester M13 9PT, UKFaculty of Biology, Medicine and Health, School of Biological Sciences, The University of Manchester, Manchester M13 9PT, UKAdenosine triphosphate-binding cassette (ABC) transporters form a ubiquitous superfamily of integral membrane proteins involved in the translocation of substrates across membranes. Human ABC transporters are closely linked to the pathogenesis of diseases such as cancer, metabolic diseases, and Alzheimer’s disease. In this study, four ABC transporters were chosen based on (I) their importance in humans and (II) their score in a structural bioinformatics screen aimed at the prediction of crystallisation propensity. The top-scoring ABC transporters’ orthologs (<i>Mus musculus</i>—mouse ABCB5, <i>Ailuropoda melanoleuca</i>—giant panda ABCB6, <i>Myotis lucifugus</i>—little brown bat ABCG1 and <i>Mus musculus</i> ABCG4) were then expressed in <i>Saccharomyces cerevisiae</i> with a combined green fluorescent protein and polyhistidine tag, enabling visualisation and purification. After partial purification and in the presence of the detergent (n-dodecyl-β-D-maltoside), the kinetic parameters of the ATP hydrolysis reactions of the orthologs were determined, as well as the extent of stimulation of their activity when presented with putative substrates. We discuss the efficiency of such bioinformatics approaches and make suggestions for their improvement and wider application in membrane protein-structure determination.https://www.mdpi.com/2077-0375/15/1/20ABC transporterortholog<i>Saccharomyces cerevisiae</i>bioinformaticsmembrane protein |
| spellingShingle | Iqra Younus Robert C. Ford Stephen M. Prince A Structural Bioinformatics-Guided Study of Adenosine Triphosphate-Binding Cassette (ABC) Transporters and Their Substrates Membranes ABC transporter ortholog <i>Saccharomyces cerevisiae</i> bioinformatics membrane protein |
| title | A Structural Bioinformatics-Guided Study of Adenosine Triphosphate-Binding Cassette (ABC) Transporters and Their Substrates |
| title_full | A Structural Bioinformatics-Guided Study of Adenosine Triphosphate-Binding Cassette (ABC) Transporters and Their Substrates |
| title_fullStr | A Structural Bioinformatics-Guided Study of Adenosine Triphosphate-Binding Cassette (ABC) Transporters and Their Substrates |
| title_full_unstemmed | A Structural Bioinformatics-Guided Study of Adenosine Triphosphate-Binding Cassette (ABC) Transporters and Their Substrates |
| title_short | A Structural Bioinformatics-Guided Study of Adenosine Triphosphate-Binding Cassette (ABC) Transporters and Their Substrates |
| title_sort | structural bioinformatics guided study of adenosine triphosphate binding cassette abc transporters and their substrates |
| topic | ABC transporter ortholog <i>Saccharomyces cerevisiae</i> bioinformatics membrane protein |
| url | https://www.mdpi.com/2077-0375/15/1/20 |
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