| http://purl.uniprot.org/citations/17848100 | http://www.w3.org/1999/02/22-rdf-syntax-ns#type | http://purl.uniprot.org/core/Journal_Citation |
| http://purl.uniprot.org/citations/17848100 | http://www.w3.org/1999/02/22-rdf-syntax-ns#type | http://purl.uniprot.org/core/Journal_Citation |
| http://purl.uniprot.org/citations/17848100 | http://www.w3.org/1999/02/22-rdf-syntax-ns#type | http://purl.uniprot.org/core/Citation |
| http://purl.uniprot.org/citations/17848100 | http://www.w3.org/2000/01/rdf-schema#comment | "Mycobacterium tuberculosis and many other members of the Actinomycetes family produce mycothiol, i.e., 1-d-myo-inosityl-2-(N-acetyl-l-cysteinyl)amido-2-deoxy-alpha-d-glucopyranoside (MSH or AcCys-GlcN-Ins), to act against oxidative and antibiotic stress. The biosynthesis of MSH is essential for cell growth and has been proposed to proceed via a biosynthetic pathway involving four key enzymes, MshA-MshD. The MSH biosynthetic enzymes present potential targets for inhibitor design. With this as a long-term goal, we have carried out a kinetic and mechanistic characterization, using steady-state and pre-steady-state approaches, of the recombinant Mycobacterium smegmatis MshC. MshC catalyzes the ATP-dependent condensation of GlcN-Ins and cysteine to form Cys-GlcN-Ins. Initial velocity and inhibition studies show that the steady-state kinetic mechanism of MshC is a Bi Uni Uni Bi Ping Pong mechanism, with ATP binding followed by cysteine binding, release of PPi, binding of GlcN-Ins, followed by the release of Cys-GlcN-Ins and AMP. The steady-state kinetic parameters were determined to be kcat equal to 3.15 s-1, and Km values of 1.8, 0.1, and 0.16 mM for ATP, cysteine, and GlcN-Ins, respectively. A stable bisubstrate analogue, 5'-O-[N-(l-cysteinyl)sulfamonyl]adenosine, exhibits competitive inhibition versus ATP and noncompetitive inhibition versus cysteine, with an inhibition constant of approximately 306 nM versus ATP. Single-turnover reactions of the first and second half reactions were determined using rapid-quench techniques, giving rates of approximately 9.4 and approximately 5.2 s-1, respectively, consistent with the cysteinyl adenylate being a kinetically competent intermediate in the reaction by MshC."xsd:string |
| http://purl.uniprot.org/citations/17848100 | http://purl.org/dc/terms/identifier | "doi:10.1021/bi7011492"xsd:string |
| http://purl.uniprot.org/citations/17848100 | http://purl.org/dc/terms/identifier | "doi:10.1021/bi7011492"xsd:string |
| http://purl.uniprot.org/citations/17848100 | http://purl.uniprot.org/core/author | "Blanchard J.S."xsd:string |
| http://purl.uniprot.org/citations/17848100 | http://purl.uniprot.org/core/author | "Blanchard J.S."xsd:string |
| http://purl.uniprot.org/citations/17848100 | http://purl.uniprot.org/core/author | "Fan F."xsd:string |
| http://purl.uniprot.org/citations/17848100 | http://purl.uniprot.org/core/author | "Fan F."xsd:string |
| http://purl.uniprot.org/citations/17848100 | http://purl.uniprot.org/core/author | "Painter G.F."xsd:string |
| http://purl.uniprot.org/citations/17848100 | http://purl.uniprot.org/core/author | "Painter G.F."xsd:string |
| http://purl.uniprot.org/citations/17848100 | http://purl.uniprot.org/core/author | "Luxenburger A."xsd:string |
| http://purl.uniprot.org/citations/17848100 | http://purl.uniprot.org/core/author | "Luxenburger A."xsd:string |
| http://purl.uniprot.org/citations/17848100 | http://purl.uniprot.org/core/date | "2007"xsd:gYear |
| http://purl.uniprot.org/citations/17848100 | http://purl.uniprot.org/core/date | "2007"xsd:gYear |
| http://purl.uniprot.org/citations/17848100 | http://purl.uniprot.org/core/name | "Biochemistry"xsd:string |
| http://purl.uniprot.org/citations/17848100 | http://purl.uniprot.org/core/name | "Biochemistry"xsd:string |
| http://purl.uniprot.org/citations/17848100 | http://purl.uniprot.org/core/pages | "11421-11429"xsd:string |
| http://purl.uniprot.org/citations/17848100 | http://purl.uniprot.org/core/pages | "11421-11429"xsd:string |
| http://purl.uniprot.org/citations/17848100 | http://purl.uniprot.org/core/title | "Steady-state and pre-steady-state kinetic analysis of Mycobacterium smegmatis cysteine ligase (MshC)."xsd:string |
| http://purl.uniprot.org/citations/17848100 | http://purl.uniprot.org/core/title | "Steady-state and pre-steady-state kinetic analysis of Mycobacterium smegmatis cysteine ligase (MshC)."xsd:string |
| http://purl.uniprot.org/citations/17848100 | http://purl.uniprot.org/core/volume | "46"xsd:string |
| http://purl.uniprot.org/citations/17848100 | http://purl.uniprot.org/core/volume | "46"xsd:string |
| http://purl.uniprot.org/citations/17848100 | http://www.w3.org/2004/02/skos/core#exactMatch | http://purl.uniprot.org/pubmed/17848100 |