Discovery of new ligand binding pathways in myoglobin by random mutagenesis.
A random library of single amino acid mutants of myoglobin was generated using a highly efficient, single-base-misincorporation random mutagenesis method to discover new ligand-binding pathways in myoglobin. A surprisingly large fraction of the library exhibits ligand-binding kinetics that are substantially different from the wild-type protein. In addition to residues 45, 64 and 68, which comprise the best studied ligand-binding pathway single mutations of several other clusters of residues far away from that pathway are discovered which profoundly affect the ligand-binding kinetics. These results provide a new approach to explore the relationship between the fluctuations in protein structure and function.
Huang
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Department of Chemistry, Stanford University, California 94305-5080, USA.
Boxer
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D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D005821:Genetic Techniques; D007700:Kinetics; D008024:Ligands; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009211:Myoglobin / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D014907:Whales
D008024:Ligands; D009211:Myoglobin