| Autor: |
Hirasawa M; Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, TX, 79409-1061, USA., Solis J; Center for Biotechnology and Genomics, Texas Tech University, Lubbock, TX, 79409-3132, USA.; Immunology and Molecular Microbiology, Texas Tech University Health Science Center, Lubbock, TX, 79430-6591, USA., Vaidyanathan N; Center for Biotechnology and Genomics, Texas Tech University, Lubbock, TX, 79409-3132, USA.; Depuy Synthes Companies, 1302 Wrights Lane East, West Chester, PA, 19380, USA., Srivastava AP; Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, TX, 79409-1061, USA.; Department of Biochemistry and Molecular Biology, Rosalind Franklin University of Medicine and Science, North Chicago, Illinoi, 60064, USA., Wynn RM; Departments of Internal Medicine and Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX, 75390-9038, USA. richard.wynn@utsouthwestern.edu., Sutton RB; Department of Cell Physiology and Molecular Biophysics, Texas Tech Health Science Center, Lubbock, TX, 79430-6551, USA., Knaff DB; Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, TX, 79409-1061, USA.; Center for Biotechnology and Genomics, Texas Tech University, Lubbock, TX, 79409-3132, USA. |
| Abstrakt: |
Based on in silico docking methods, five amino acids in glutamate synthase (Gln-467, His-1144, Asn-1147, Arg-1162, and Trp-676) likely constitute key binding residues in the interface of a glutamate synthase:ferredoxin complex. Although all interfacial mutants studied showed the ability to form a complex under low ionic strength, these docking mutations showed significantly less ferredoxin-dependent activities, while still retaining enzymatic activity. Furthermore, isothermal titration calorimetry showed a possible 1:2 molar ratio between the wild-type glutamate synthase and ferredoxin. However, each of our interfacial mutants showed only a 1:1 complex with ferredoxin, suggesting that the mutations directly affect the glutamate synthase:ferredoxin heterodimer interface. |
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