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

Date
2014-01-15
Authors
Chang, Melvin
Contributor
Advisor
Lenny, James
Department
Biology
Instructor
Depositor
Speaker
Researcher
Consultant
Interviewer
Journal Title
Journal ISSN
Volume Title
Publisher
University of Hawaii at Manoa
Volume
Number/Issue
Starting Page
Ending Page
Alternative Title
Abstract
A 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.
Description
Keywords
Citation
Extent
viii, 47 pages
Format
Geographic Location
Time Period
Related To
Rights
All 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.
Rights Holder
Email libraryada-l@lists.hawaii.edu if you need this content in ADA-compliant format.