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10.31 Metalloproteases are enzymes that require a metalion as a cofactor when they cleave peptide bonds. Members of one family of metalloproteases share the following consensus amino acid sequence in their catalytic site:His-Glu-X-Gly-His-Asp-X-Gly-X-X-His-Asp (X is any amino acid). Structural models of the catalytic site developed from X-ray crystallographic data suggest that the second amino acid, glutamate, is essential for proteolytic activity.Outline the experimental steps you would take to test this hypothesis. Assume you possess a cDNA encoding a metalloprotease having the consensus sequence, and canmeasure metalloprotease activity in a biochemical assay.
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