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Bovine pancreatic ribonuclease A (RNase A) catalyzes the cleavage of the P-O 5′ bond in RNA. Although this enzyme has been the object of much seminal work in biological chemistry, the nature of its rate-limiting transition state and its catalytic rate enhancement had been unknown. Here, the value of k cat /K m for the cleavage of UpA by wild-type RNase A was found to be inversely...