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Article
Nature Structural Biology  1, 226 - 229 (1994)
doi:10.1038/nsb0494-226

Discovery of new ligand binding pathways in myoglobin by random mutagenesis

Xiaohua Huang1 & Steven G. Boxer1

1Department of Chemistry, Stanford University, Stanford, California 94305-5080, USA

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.

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