BBa_K314986 1 thisisthen -- No description -- thisisthenicknamethisisthenicknamethisisthenicknamethisisthenicknamethisisthen 2010-10-02T11:00:00Z 2015-05-08T01:11:55Z To build the mutant proteins, we follow the path of the central dogma. First, we created DNA that contains our mutations. Second, we induced our transformed cells containing the desired DNA to express the mutant proteins. Lastly, we harvested the proteins by lysing open the cells and filtering out non-desired cell components. There are two main obstacles limiting natural CapD as an Anthrax therapeutic. First, natural CapD is a difficult to express dimer, requiring an auto-cleavage to activate. Second, CapD is a better transpeptidase than poly-????-D-glutamate hydrolase, limiting its Anthrax decapsulating potential. To solve the first problem, we created a circular permuted, monomeric version of CapD that is easy to express and quantify. To improve hydrolysis, we used FoldIt, a computational toolbox, to design active site mutations aimed to increase hydrolysis over transpeptidation. true false _496_ 0 6395 9 Discontinued false To increase the hydrolytic ability of CapD_CP, we made point mutations to the active site. We focused our attention on two types of mutations. First, we created point mutations that can establish hydrogen bondings to a modeled transition state of our substrate in an attempt to lower the activation energy, making hydrolysis more favorable. Second, we mutated the active site into a more open and polar area in an attempt to increase the ease with which water can enter and participate in a hydrolysis reaction. false annotation2084281 1 BBa_K314987 range2084281 1 1 144 annotation2084280 1 s2 range2084280 1 75 90 annotation2084282 1 mypr range2084282 1 154 164 annotation2084297 1 s2 range2084297 1 379 394 annotation2084288 1 mypr range2084288 1 306 316 annotation2084296 1 s2 range2084296 1 379 394 annotation2084284 1 BBa_K314988 range2084284 1 153 296 annotation2084283 1 mypr range2084283 1 154 164 annotation2084279 1 s1 range2084279 1 60 65 annotation2084287 1 mypr range2084287 1 306 316 annotation2084290 1 rib1 range2084290 1 318 330 annotation2084278 1 capd range2084278 1 30 55 annotation2084285 1 mypr range2084285 1 306 316 annotation2084294 1 s1 range2084294 1 364 369 annotation2084293 1 capd range2084293 1 334 359 annotation2084276 1 mypr range2084276 1 2 12 annotation2084289 1 mypr range2084289 1 306 316 annotation2084292 1 capd range2084292 1 334 359 annotation2084295 1 s1 range2084295 1 364 369 annotation2084286 1 mypr range2084286 1 306 316 annotation2084291 1 rib1 range2084291 1 318 330 annotation2084298 1 BBa_K314989 range2084298 1 305 448 annotation2084277 1 rib1 range2084277 1 14 26 BBa_K314986_sequence 1 ttttcgggctcgcgtctgagagggaaaggctgcggctaggcatacgtactgacgatcgtacgcggcgctcccttttcgggctcgcgtctgagagggaaaggctgcggctaggcatacgtactgacgatcgtacgcggcgctccctactagagttttcgggctcgcgtctgagagggaaaggctgcggctaggcatacgtactgacgatcgtacgcggcgctcccttttcgggctcgcgtctgagagggaaaggctgcggctaggcatacgtactgacgatcgtacgcggcgctccctactagagttttcgggctcgcgtctgagagggaaaggctgcggctaggcatacgtactgacgatcgtacgcggcgctcccttttcgggctcgcgtctgagagggaaaggctgcggctaggcatacgtactgacgatcgtacgcggcgctccc igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 James Alastair McLaughlin Chris J. Myers 2017-03-06T15:00:00.000Z