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dc.contributor.advisorNinfa, Alexander J.
dc.contributor.authorOuillette, Ryan Matthew
dc.descriptioniv, 33 p.en_US
dc.description.abstractUTase/UR is a key component of the bicyclic cascade transduction pathway that regulates nitrogen assimilation in Escherichia coli via the protein glutamine synthetase (GS). This study examined the effects that specific site-directed mutations of highly conserved residues in the HD (phosphohydrolase) and ACT domains of the uridylyltransferase/ uridylyl-removing enzyme (UTase/UR) had on protein function. The effect mutations had on UTase/UR mutants was examined by means of a GS assay, in which UTase/UR behavior was extrapolated from the activity level of GS. An attempt was also made to purify one version of the mutant protein along with the wild-type. We discovered that mutations of these highly conserved domains suggest a loss or reduction of the uridylyl-removing activity of UTase/UR, but the protein maintained its uridylyl-transferase ability. Also, our d~ta suggest that the UTase/UR protein cannot be purified using a polyhistidine tag and Ni-NTA column approach due to possible problems with protein folding. Further investigations should be carried out to determine how the mutant UTase/UR enzyme loses its uridylyl-removing ability and what prevents it from being able to be purified using a His-tag.en_US
dc.description.sponsorshipDepartment of Biological Chemistry. University of Michigan. Ann Arbor, Michigan.
dc.publisherKalamazoo Collegeen_US
dc.relation.ispartofKalamazoo College Biology Senior Individualized Projects Collection
dc.relation.ispartofseriesSenior Individualized Projects. Biology;
dc.rightsU.S. copyright laws protect this material. Commercial use or distribution of this material is not permitted without prior written permission of the copyright holder.
dc.titleFunctional Analysis of the HD and ACT domains of the E. coli signal-transducing Uridylyltransferase/Uridylyl-removing enzyme (UTaselUR)en_US
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  • Biology Senior Individualized Projects [1489]
    This collection includes Senior Individualized Projects (SIP's) completed in the Biology Department. Abstracts are generally available to the public, but PDF files are available only to current Kalamazoo College students, faculty, and staff.

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