The Isolation And Characterization Of A Yeast Protein Which Specifically Inhibits Yeast Proteinase A

dc.contributor.advisorLenny, James
dc.contributor.authorChang, Melvin
dc.contributor.departmentBiology
dc.date.accessioned2014-01-15T20:05:35Z
dc.date.available2014-01-15T20:05:35Z
dc.date.issued2014-01-15
dc.description.abstractA yeast protein was discovered which specifically inhibits proteinase A of Saccharomyces cerevisiae. The inhibitor was purified 8.4-fold using boiling, dialysis and DEAE-cellulose ion-exchange chromatographic steps. The inhibitor, IA, was found to be an unusually thermostable, nondialyzable protein which was not precipitable with trichloroacetic acid, and which adsorbed to DEAE-cellulose at pH 9-10, and CM-Sephadex at pH 6.0. Elution of the inhibitor from DEAE – cellulose was accomplished with 0.01 M Borax buffer, pH 9.1, containing 0.05 M NaCl. Sephadex G-75 gel-filtration and SDS molecular weight disc electrophoresis suggest a molecular weight of about 13,000 for IA. Digestion by trypsin, chymotrypsin, pepsin, subtilisin and yeast carboxypeptidase demonstrated the inhibitor to be of protein nature. IA was found in the greatest concentration in the cytosol of the cell. Possible digestion of IA at pH 2.5 by proteinase A was observed. Digestion of the inhibitor by yeast proteases at pH 6.8 was clearly demonstrated. Lenney's report (6) of proteinase A activation in pH 5.0, yeast autolysates was also confirmed.
dc.format.extentviii, 47 pages
dc.identifier.urihttp://hdl.handle.net/10125/32008
dc.publisherUniversity of Hawaii at Manoa
dc.rightsAll UHM Honors Projects are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission from the copyright owner.
dc.titleThe Isolation And Characterization Of A Yeast Protein Which Specifically Inhibits Yeast Proteinase A
dc.typeTerm Project
dc.type.dcmiText

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