Characterization of the cleavage specificity of a subtilisin‐like serine proteinase from Ophiostoma piceae by liquid chromatography/mass spectrometry and tandem MS

FEBS Letters

Chow, David T.²  Abraham, Linda D.¹  Breuil, Colette¹ 

[1] Chair of Forest Products Biotechnology, Department of Wood Science, Faculty of Forestry, University of British Columbia, Vancouver, B.C. V6T 1Z3, Canada;Biomedical Research Centre, University of British Columbia, Vancouver, B.C. V6T 1Z3, Canada

关键词: Electrospray ionization mass spectrometry;    Tandem mass spectrometry;    Subtilisins;    Serine proteinases;    Sapstain;    Ophiostoma;   

DOI  :  10.1016/0014-5793(95)01107-P

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A proteinase secreted by the sapstaining fungus Ophiostoma piceae is thought to be necessary for the primary retrieval of nitrogen from wood proteins. By using mass spectrometry (MS) techniques, we have estabilished the cleavage specificity of this subtilisin-like serine proteinase. This work demonstrated the potential of MS in determining cleavage specificities of newly isolated proteinases in a relatively short time frame, and determined that the O. piceae proteinase showed a substrate specificity similar to that of proteinase K. Primary cleavage of the insulin B-chain occurred between Leu¹⁵ and Tyr¹⁶. In addition numerous secondary cleavage sites occurred after hydrophobic, polar, and charged amino acids indicating a broad specificity.

【 授权许可】

Unknown