Steitz was awarded the 2009 Nobel Prize in Chemistry along with Venkatraman Ramakrishnan and Ada Yonath "for studies of the structure and function of the ribosome". Steitz also won the Gairdner International Award in 2007 "for his studies on the structure and function of the ribosome which showed that the peptidyl transferase was an RNA catalyzed reaction, and for revealing the mechanism of inhibition of this function by antibiotics".
Born in Milwaukee, Wisconsin, Steitz studied chemistry as an undergraduate at Lawrence University in Appleton, Wisconsin, graduating in 1962. While there he was a member of the fraternity, Delta Tau Delta, the Delta Nu chapter. In June 2010, the University renamed its chemistry building Thomas A. Steitz Hall of Science.
He received a Ph.D. in biochemistry and molecular biology from Harvard University in 1966 where he worked under the direction of subsequent 1976 chemistry Nobel Prize winner William N. Lipscomb, Jr.
While at Harvard, after the training task of determining the structure of the small molecule methyl ethylene phosphate, Steitz made contributions to determining the atomic structures of carboxypeptidase A and aspartate carbamoyltransferase, each the largest atomic structure determined in its time.

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