Department of Biochemistry, Eotvos Lorand University, and 2Biotechnology Research Group of the Hungarian Academy of Sciences, ´ ´ ´ ´ Pazmany setany 1/C, Budapest, Hungary The molecular mechanism of the autolysis of rat a-chymotrypsin B was investigated. In addition to the two already known autolytic sites, Tyr146 and Asn147, a new site formed by Phe114 was identified. The former two sites and the latter one are located in the autolysis and the interdomain loops, respectivel