Site-Specific Mutation of Nitrogenase Proteins

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Nitrogen fixation component enzymes have been isolated and crystallized. The 3-dimensional structures will soon be known. The objective of this award is to the methods of side specific mutagenesis to augment biochemical and biophysical studies of the Fe-protein component. The experiments are designed to elucidate: a) the role of the five conserved cysteines, and arginine-100 and b) the amino acid residues involved in complex formation between components of nitrogenase, flavodoxin binding and ATP binding. The effects of substitutions will be studied by biochemical, biophysical and crystallographic methods. The results will enhance our understanding of the structure and function of nitrogenase, an enzyme which converts molecular nitrogen into ammonia and makes it utilizable by all the forms of life on earth.