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Expression and Purification of Murine Tripeptidyl Peptidase II
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Biochemistry and Microbiology.
2012 (Swedish)Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
Abstract [en]

Tripeptidyl peptidase II (TPPII) is an exopeptidase which cleaves tripeptides from theN-terminus of peptides. The exact functional role of TPPII is still a matter of investigation. Itis believed that the enzyme is primarily involved in intracellular protein degradation, where itcooperates with the proteasome and other peptidases to degrade proteins into free aminoacids. These amino acids can subsequently be used in the production of new proteins. The aimof this work was to express murine wild type TPPII using E. coli and thereafter purify theenzyme from the bacterial lysate. Methods used for the purification included protein andnucleic acid precipitation, anion exchange chromatography, hydrophobic interactionchromatography and gel filtration. The presence of TPPII was determined using activityassay, western blot and SDS-PAGE. Despite the fact that some modification is still needed,the purification yielded a total of 34μg TPPII with a purity of approximately 60%. Thispurified enzyme can be used for future functional characterization.

Place, publisher, year, edition, pages
2012. , 33 p.
Keyword [en]
E. coli expression, Anion exchange chromatography, Hydrophobic interaction chromatography, Gel filtration, Steady state kinetics
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:uu:diva-177009OAI: oai:DiVA.org:uu-177009DiVA: diva2:538742
Educational program
Biomedicinska analytikerprogrammet
Uppsok
Medicine
Supervisors
Examiners
Available from: 2012-07-23 Created: 2012-07-02 Last updated: 2012-07-23Bibliographically approved

Open Access in DiVA

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