Structural Studies on PP2A and Methods in Protein Production
Stockholm University, Faculty of Science, Department of Biochemistry and Biophysics2008 (English)Doctoral thesis, comprehensive summary (Other academic)
PP2A is a major phosphatase in the cell that participates in multiple cell signaling pathways. It is a heterotrimer of a core dimer and variable regulatory subunits. Details of its structure, function and regulation are slowly emerging. Here, the structure of two regulators of PP2A are de-scribed; PTPA and B56γ. PTPA is a highly conserved enzyme that plays a crucial role in PP2A activity but whose biochemical function is still unclear. B56γ is a PP2A regulatory subunit linked to cancer and the structure presented here of B56γ in its free form is particularly valuable in light of the recent structures of the PP2A holoenzyme and core dimer.
Protein production is a major bottleneck in structural genomic projects. Here, we describe two novel methods for improved protein production. The first is a colony based screening method where any DNA library can be screened for soluble expression of recombinant proteins in E.coli. The second method involves improvements of the well established IMAC purification method. We have seen that a low molecular weight component of E.coli lysate decreases the binding capacity of IMAC columns and by removing the low molecular weight components, recombinant proteins only present at low levels in E.coli lysate can be purified, which has previously been believed to be unfeasible.
Place, publisher, year, edition, pages
Stockholm: Institutionen för biokemi och biofysik , 2008. , 53 p.
PP2A, PTPA, B56γ, Protein Production
Research subject Biochemistry
IdentifiersURN: urn:nbn:se:su:diva-8261ISBN: 978-91-7155-750OAI: oai:DiVA.org:su-8261DiVA: diva2:199933
2008-11-14, Magnélisalen, Kemiska övningslaboratoriet, Svante Arrhenius väg 12 A, Stockholm, 10:00
Van Tilbeurgh, Herman, Professor
Nordlund, Pär, Professor
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