[{"key":"dc.contributor.author","value":"Moore, Todd Robert","language":null},{"key":"dc.date.accessioned","value":"2009-08-05T19:08:59Z","language":null},{"key":"dc.date.available","value":"2009-08-05T19:08:59Z","language":null},{"key":"dc.date.issued","value":"2001","language":null},{"key":"dc.identifier.uri","value":"http:\/\/hdl.handle.net\/2429\/11743","language":null},{"key":"dc.description.abstract","value":"The matrix metalloproteinase (MMP) family of endopeptidases can collectively\r\ndegrade many components of the extracellular matrix. Their proteolytic activities\r\nhave been implicated in normal processes such as extracellular matrix turnover\r\nand certain pathological conditions such as periodontitis, arthritis, and tumour\r\nmetastasis.\r\nBased upon domain composition gelatinase A is separated from the other MMPs\r\nby the insertion of three contiguous fibronectin type II modules into its catalytic\r\ndomain. Since it was determined that a recombinant domain consisting of the\r\nthree fibronectin type II modules bound to native type I collagen this domain was\r\ntermed the collagen binding domain (CBD). The function of the CBD is to donate\r\nsubstrate binding exosites to allow for broader enzyme specificity. Considering\r\nthat the CBD contains exosites for binding native\/denatured collagen, this\r\ndomain was subjected to site-directed mutagenic studies to elucidate essential\r\nresidues involved in collagen binding.\r\nThe binding properties of the collagen binding domain to denatured type I\r\ncollagen (gelatin) was investigated using a recombinant protein constructed of\r\nthe second and third fibronectin type II module (rCBD23). Ten mutations were\r\nperformed within the rCBD23 protein. Since the binding of substrate to the CBD\r\nis via a hydrophobic pocket, the mutations F264A, F264Y, F266A, F266Y,\r\nF322A, F324A, F322Y, F324Y, F264A\/F322A, and F266A\/F324A were\r\nintroduced into rCBD23 in order to determine the effect of removing hydrophobic\r\ncharacter.\r\nIt was found that there was a decrease in the binding of the mutant proteins that\r\nwas proportional to the distance between the mutated residue side chain and a\r\nstrictly conserved tryptophan at the base of the hydrophobic pocket. Complete\r\nabrogation of gelatin binding was observed in the double mutant F266A\/F324A.\r\nThe observed effect was proposed to be the result of destabilisation of the strictly\r\nconserved tryptophan that forms the base of the hydrophobic pocket.\r\nThese studies have furthered our knowledge and understanding of the\r\ninteractions of CBD with substrate. The present study, combined with results\r\nfrom other studies, could be used to synthesize compounds that are specific to\r\nthe CBD and are able to irreversibly inhibit the binding of CBD to substrate. A\r\ndrug that would preferentially block binding of the CBD to collagen is proposed to\r\nreduced the metastasis of tumour cells by abolishment of type IV collagen\r\nbinding. The overall effect is decreased tumour migration from the primary site of\r\ndevelopment and therefore substantially decreasing tumour progression.","language":"en"},{"key":"dc.format.extent","value":"4601806 bytes","language":null},{"key":"dc.format.mimetype","value":"application\/pdf","language":null},{"key":"dc.language.iso","value":"eng","language":"en"},{"key":"dc.publisher","value":"University of British Columbia","language":null},{"key":"dc.rights","value":"For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https:\/\/open.library.ubc.ca\/terms_of_use.","language":null},{"key":"dc.title","value":"Characterisation of the collagen binding domain of gelatinase A : involvement of specific residues in the fibronectin type II modules in substrate recognition","language":"en"},{"key":"dc.type","value":"Text","language":null},{"key":"dc.degree.name","value":"Master of Science - MSc","language":"en"},{"key":"dc.degree.discipline","value":"Dental Science","language":null},{"key":"dc.degree.grantor","value":"University of British Columbia","language":null},{"key":"dc.date.graduation","value":"2001-05","language":"en"},{"key":"dc.type.text","value":"Thesis\/Dissertation","language":"en"},{"key":"dc.description.affiliation","value":"Dentistry, Faculty of","language":null},{"key":"dc.degree.campus","value":"UBCV","language":"en"},{"key":"dc.description.scholarlevel","value":"Graduate","language":"en"}]