Purification of a novel type of SDS-dependent protease in maize [Zea mays] using a monoclonal antibody
A protease which was activated by SDS was purified to homogeneity from maize laves. On the basis of its proteolytic activity towards ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) or a synthesized peptide, the purification was carried out using immunoaffinity chromatography with a monoclo...
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Published in | Plant and cell physiology Vol. 39; no. 1; pp. 106 - 114 |
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Main Authors | , , , , , , , |
Format | Journal Article |
Language | English |
Published |
Oxford
Oxford University Press
01.01.1998
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Subjects | |
Online Access | Get full text |
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Summary: | A protease which was activated by SDS was purified to homogeneity from maize laves. On the basis of its proteolytic activity towards ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) or a synthesized peptide, the purification was carried out using immunoaffinity chromatography with a monoclonal antibody raised against a partially purified enzyme by native gradient PAGE. The purified protease showed three bands at 40, 15, and 13 kDa on SDS-PAGE, indicating that it was composed of heterogeneous subunits. The protease was specifically activated by SDS (optimum = 0.4% for Rubisco proteolysis), but not by poly-L-lysine, fatty acids, or ATP. The protease had a pH optimum around 4.9. beta-Mercaptoethanol stimulated the activity only in the presence of SDS. The proteolytic activity was sensitive to E-64 and leupeptin but was resistant to EDTA, suggesting that the enzyme was an SH-protease. Thus, this enzyme is a novel type of SDS-dependent protease which differs from proteasome, matrix metalloproteinase, and other proteases reported in many organisms |
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Bibliography: | 1998003338 F70 H20 istex:43B5B552C194597C70E9AC51DE5CF7FD6F77BEDB ark:/67375/HXZ-D77MH3NX-X ArticleID:39.1.106 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0032-0781 1471-9053 |
DOI: | 10.1093/oxfordjournals.pcp.a029281 |