Nuclease B. A possible precursor of nuclease A, an extracellular nuclease of Staphylococcus aureus

Abstract

During purification of nuclease (redisignated as nuclease A in the present studies) from the culture media of Staphylococcus aureus strain Foggi, three enzymacally active second species (nucleases B1, B2, and B3) were isolated as a mixture by ion exchange chromatography. Examination of the amino acid sequence of these second species indicates that nucleases B1, B2, and B3 apparently contain the same sequence as that of nuclease A with an extra sequence Ser-Gln-Thr-Asp-Asx-Gly-Val-Asx-Arg-Ser-Gly-Ser-Glu-Asp-Pro-Thr-Val-Tyr-Ser linked through a peptide bond to the NH2 terminus of the nuclease A portion. In nuclease B1 the 2 residues indicated by Asx are aspartic acid, in nuclease B2 the first and the second Asx from the NH2 terminus are aspartic acid and asparagine, respectively, and in nuclease B3 both Asx are asparagine. These second species do not contain a significant amount of carbohydrate. The extra amino acid sequence appears to be flexible and does not interfere with the ordered structure and function of the nuclease A portion. The nuclease A portion was recovered, in part, from a mixture of these nuclease B species after digestion with staphylococcal protease in the presence of ligands, deoxythymidine 3',5'-diphosphate, and calcium ion. Thus, these nuclease B species may be closely related to, if not identical with, a precursor of nuclease A. Similar second species of nuclease have been found in strain V8.