Complete cysteine-scanning mutagenesis of the Salmonella typhimurium melibiose permease
| Autor: | Elena B. Tikhonova, Lan Guan, Aaron C. Scarpa, Parameswaran Hariharan, Kelsey J. Markham, Satoshi Katsube |
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| Rok vydání: | 2021 |
| Předmět: |
Models
Molecular Salmonella typhimurium MFS transporter Stereochemistry Mutant Biological Transport Active Biochemistry Molecular recognition α-NPG α-nitrophenyl galactoside Bacterial Proteins structure and function analysis Cysteine Molecular Biology DDMB dodecyl melibioside Binding Sites Ion Transport Symporters biology Cys-less 4 endogenous Cys was replaced by Ala Chemistry MelBSt MelBSt the melibiose permease of Salmonella typhimurium Sodium Mutagenesis MFS major facilitator superfamily Transporter Cell Biology biology.organism_classification cation-coupled symporter Major facilitator superfamily Cys-scanning mutagenesis DW2 strain melA+ ΔmelB ΔlacZY MelBEc the melibiose permease of E. coli Symporter Cys-scanning mutagenesis Cysteine-scanning mutagenesis Bacteria Research Article |
| Zdroj: | The Journal of Biological Chemistry |
| ISSN: | 0021-9258 |
| Popis: | The melibiose permease of Salmonella typhimurium (MelBSt) catalyzes the stoichiometric symport of galactopyranoside with a cation (H+, Li+, or Na+) and is a prototype for Na+-coupled major facilitator superfamily (MFS) transporters presenting from bacteria to mammals. X-ray crystal structures of MelBSt have revealed the molecular recognition mechanism for sugar binding; however, understanding of the cation site and symport mechanism is still vague. To further investigate the transport mechanism and conformational dynamics of MelBSt, we generated a complete single-Cys library containing 476 unique mutants by placing a Cys at each position on a functional Cys-less background. Surprisingly, 105 mutants (22%) exhibit poor transport activities ( |
| Databáze: | OpenAIRE |
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