2p2n: Difference between revisions
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==Overview== | ==Overview== | ||
AnsA is the cytoplasmic asparaginase from Escherichia coli involved in | AnsA is the cytoplasmic asparaginase from Escherichia coli involved in intracellular asparagine utilization. Analytical ultracentifugation and X-ray crystallography reveal that AnsA forms a tetrameric structure as a dimer of two intimate dimers. Kinetic analysis of the enzyme reveals that AnsA is positively cooperative, displaying a sigmoidal substrate dependence curve with an [S](0.5) of 1 mM L-asparagine and a Hill coefficient (n(H)) of 2.6. Binding of L-asparagine to an allosteric site was observed in the crystal structure concomitant with a reorganization of the quarternary structure, relative to the apo enzyme. The carboxyl group of the bound asparagine makes salt bridges and hydrogen bonds to Arg240, while the N(delta2) nitrogen interacts with Thr162. Mutation of Arg240 to Ala increases the [S](0.5) value to 5.9 mM, presumably by reducing the affinity of the site for L-asparagine, although the enzyme retains cooperativity. Mutation of Thr162 to Ala results in an active enzyme with no cooperativity. Transmission of the signal from the allosteric site to the active site appears to involve subtle interactions at the dimer-dimer interface and relocation of Gln118 into the vicinity of the active site to position the probable catalytic water molecule. These data define the structural basis for the cooperative regulation of the intracellular asparaginase that is required for proper functioning within the cell. | ||
==About this Structure== | ==About this Structure== | ||
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==Reference== | ==Reference== | ||
Crystal | Crystal structure and allosteric regulation of the cytoplasmic Escherichia coli L-asparaginase I., Yun MK, Nourse A, White SW, Rock CO, Heath RJ, J Mol Biol. 2007 Jun 8;369(3):794-811. Epub 2007 Mar 30. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=17451745 17451745] | ||
[[Category: Asparaginase]] | [[Category: Asparaginase]] | ||
[[Category: Escherichia coli]] | [[Category: Escherichia coli]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Heath, R | [[Category: Heath, R J.]] | ||
[[Category: Nourse, A.]] | [[Category: Nourse, A.]] | ||
[[Category: Rock, C | [[Category: Rock, C O.]] | ||
[[Category: White, S | [[Category: White, S W.]] | ||
[[Category: Yun, M | [[Category: Yun, M K.]] | ||
[[Category: ASN]] | [[Category: ASN]] | ||
[[Category: ASP]] | [[Category: ASP]] | ||
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[[Category: asparaginase]] | [[Category: asparaginase]] | ||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 18:25:11 2008'' | ||
Revision as of 19:25, 21 February 2008
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Crystal Structure and Allosteric Regulation of the Cytoplasmic Escherichia coli L-Asparaginase I
OverviewOverview
AnsA is the cytoplasmic asparaginase from Escherichia coli involved in intracellular asparagine utilization. Analytical ultracentifugation and X-ray crystallography reveal that AnsA forms a tetrameric structure as a dimer of two intimate dimers. Kinetic analysis of the enzyme reveals that AnsA is positively cooperative, displaying a sigmoidal substrate dependence curve with an [S](0.5) of 1 mM L-asparagine and a Hill coefficient (n(H)) of 2.6. Binding of L-asparagine to an allosteric site was observed in the crystal structure concomitant with a reorganization of the quarternary structure, relative to the apo enzyme. The carboxyl group of the bound asparagine makes salt bridges and hydrogen bonds to Arg240, while the N(delta2) nitrogen interacts with Thr162. Mutation of Arg240 to Ala increases the [S](0.5) value to 5.9 mM, presumably by reducing the affinity of the site for L-asparagine, although the enzyme retains cooperativity. Mutation of Thr162 to Ala results in an active enzyme with no cooperativity. Transmission of the signal from the allosteric site to the active site appears to involve subtle interactions at the dimer-dimer interface and relocation of Gln118 into the vicinity of the active site to position the probable catalytic water molecule. These data define the structural basis for the cooperative regulation of the intracellular asparaginase that is required for proper functioning within the cell.
About this StructureAbout this Structure
2P2N is a Single protein structure of sequence from Escherichia coli with , , and as ligands. Active as Asparaginase, with EC number 3.5.1.1 Full crystallographic information is available from OCA.
ReferenceReference
Crystal structure and allosteric regulation of the cytoplasmic Escherichia coli L-asparaginase I., Yun MK, Nourse A, White SW, Rock CO, Heath RJ, J Mol Biol. 2007 Jun 8;369(3):794-811. Epub 2007 Mar 30. PMID:17451745
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