MacDonald, Justin A.Grassie, Michael2017-12-182017-12-182012Grassie, M. (2012). Investigating the phosphorylation of ser695/thr696 and ser852/thr853 regions of smooth muscle myosin phosphatase targeting subunit (Master's thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca. doi:10.11575/PRISM/4944http://hdl.handle.net/1880/105945Bibliography: p. 122-135A few pages are in colour.Includes copy of electronic thesis agreement. Original copy with original Partial Copyright Licence.MYPTl has proven to be a key regulator in smooth muscle contraction and relaxation through the mechanisms of Ca2+ sensitization and desensitization. The regulation of MYPTl is controlled through the phosphorylation of several key residues. Kinase assays were carried out to confirm which amino acid residues of MYPT 1 were targeted by PKA and PKG in vitro. The following residues were then examined for functional significance in vitro and in situ with the use of recombinant proteins and RT A smooth muscle. From our studies we demonstrate the first direct evidence of in situ dual phosphorylation of MYPTl at Ser695/Thr696 and Ser852/Thr853. Also, that pre­phosphorylation of Ser852 by PKA/PKG has an inhibitory effect on the ability of ROK to phosphorylate the neighboring inhibitory residue of Thr853, similar to the effect previously observed for the Ser695/Thr696 region. Furthermore, the pre-phosphorylation ofThr696 by ROK decreased the ability of PKA/PKG to phosphorylate Ser695 in vitro and in situ.xii, 136 leaves : ill. ; 30 cm.engUniversity of Calgary graduate students retain copyright ownership and moral rights for their thesis. You may use this material in any way that is permitted by the Copyright Act or through licensing that has been assigned to the document. For uses that are not allowable under copyright legislation or licensing, you are required to seek permission.master thesis10.11575/PRISM/4944