Show simple item record

dc.contributor.authorTezcan, Mehtap Yilmazen_US
dc.date.accessioned2015-08-07T04:10:01Z
dc.date.available2015-08-07T04:10:01Z
dc.date.issued2013
dc.date.submitted2013
dc.identifier.other(ALMA)contemp
dc.identifier.urihttps://hdl.handle.net/2144/12897
dc.descriptionThesis (Ph.D.)--Boston University PLEASE NOTE: Boston University Libraries did not receive an Authorization To Manage form for this thesis or dissertation. It is therefore not openly accessible, though it may be available by request. If you are the author or principal advisor of this work and would like to request open access for it, please contact us at open-help@bu.edu. Thank you.en_US
dc.description.abstractCalcium/calmodulin dependent kinase II (CaMKII) 1s a serine/threonine phosphotransferase that is capable of long-term retention of activity due to autophosphorylation at a specific threonine residue (Thr^287) within each subunit of its oligomeric structure. Using an antisense oligonucleotide, CaMKII γ was previously demonstrated to lead to decreased contractility of blood vessels. However, the mechanism of this process remains unknown. In this thesis, I show that the six CaMKII γ smooth muscle variants are differentially targeted when expressed in cultured cells. I also show that CaMKII γ variants disrupt microtubules upon Ca^2+ stimulation in a variant-independent manner. Similar microtubule disassembly was shown in A7r5 vascular smooth muscle cells which display CaMKII properties similar to blood vessels. The major discovery of this thesis is the determination of a novel and physiologically significant phosphorylation-dependent function of CaMKII γ. Phosphorylation of Ser^26 was found to switch off kinase activity. I show that phosphorylation of CaMKII gamma at Ser^26, a residue located within the ATP binding site, terminates the sustained activity of the enzyme. To test the physiological importance of phosphorylation at Ser^26, a phosphospecific Ser^26 antibody was generated and I demonstrated an increase of Ser^26 phosphorylation upon depolarization and contraction of blood vessels. To determine if the phosphorylation of Ser^26 affects the kinase activity, the Ser^26 residue was mutated into an Ala or Asp. The recombinant S26D mutant mimicking the phosphorylated state of CaMKII displays a dramatic decrease in Thr^287 autophosphorylation levels in vitro and greatly reduces the catalytic activity towards an exogenous substrate (autocamtide-3), while the S26A mutation has no effect. These data combined with molecular modeling indicate that a negative charge at Ser^26 of CaMKII gamma inhibits the catalytic activity of the enzyme towards its autophosphorylation site at Thr^287 most likely by blocking ATP binding. Here I propose that Ser^26 phosphorylation constitutes an important mechanism for switching off CaMKII activity, which may play an important role in vascular contractility.en_US
dc.language.isoen_US
dc.publisherBoston Universityen_US
dc.titlePhosphorylation dependent regulation of CaMKII gamma activity and its implication in vascular smooth muscle functionen_US
dc.typeThesis/Dissertationen_US
etd.degree.nameDoctor of Philosophyen_US
etd.degree.leveldoctoralen_US
etd.degree.disciplineMolecular, Cell Biology & Biochemistryen_US
etd.degree.grantorBoston Universityen_US


This item appears in the following Collection(s)

Show simple item record