Isolation and characterization of an enzymically active fragment of papain.
نویسندگان
چکیده
Evidence has been presented earlier (2, 3) that leucine aminopeptidase can degrade about two-thirds of the molecule of mercuripapain, liberating free amino acids, without altering the enzymic activity of the papain when it is reactivated. Although several experimental results supported this conclusion, the best evidence came from two types of observations: first, new amino end groups different from the end group of intact papain, were found in the degraded enzyme, and second, amino acid analysis of degraded mercuripapain revealed that it was different from the native enzyme only by those amino acids liberated by the aminopeptidase (3). Experiments reported in this paper provide additional evidence that mercuripapain can be degraded to an enzymically active fragment. Chromatographic procedures were developed which have allowed the separation of the active fragment in homogeneous form. The active substance contains approximately 76 residues in contrast to the 185 residues of intact papain. The fragment also differs from intact papain in chromatographic behavior, in ultraviolet absorption spectrum, amino end group, electrophoretic mobility, and molecular weight. Although the specific activity of the degraded enzyme is about 3 times that of intact papain, both kinds of enzyme have an identical substrate specificity and show similar behavior with respect to denaturation by heat, acid, or urea.
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عنوان ژورنال:
- The Journal of biological chemistry
دوره 235 شماره
صفحات -
تاریخ انتشار 1960