2dt9: Difference between revisions

Revision as of 18:02, 21 February 2008

Crystal structure of the regulatory subunit of aspartate kinase from Thermus flavus

OverviewOverview

To reveal the catalytic mechanism of Thermus aspartate kinase, each of 29 amino acid residues that were highly conserved in the sequenced aspartate kinases, was replaced with alanine or leucine by PCR site-directed mutagenesis. Comparison of the kinetic parameters of these mutants with those of the wild-type aspartate kinase suggested that Thr47 was involved in binding aspartate and that Lys7 and Glu74 were involved in catalysis. Analysis of the effective concentrations of magnesium ion on the activity showed that the mutants with replacements at Ser41, Thr47, Asp154 and Asp182 required higher concentrations of magnesium ion. This suggests that these four residues play important roles in the binding of magnesium ions which are required for enzymatic activity.

About this StructureAbout this Structure

2DT9 is a Single protein structure of sequence from Thermus thermophilus with and as ligands. Active as Aspartate kinase, with EC number 2.7.2.4 Full crystallographic information is available from OCA.

ReferenceReference

Kinetic and mutation analyses of aspartate kinase from Thermus flavus., Kobashi N, Nishiyama M, Tanokura M, J Biosci Bioeng. 1999;87(6):739-45. PMID:16232547

Page seeded by OCA on Thu Feb 21 17:02:28 2008

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 ==Overview==
-To reveal the catalytic mechanism of Thermus aspartate kinase, each of 29, amino acid residues that were highly conserved in the sequenced aspartate, kinases, was replaced with alanine or leucine by PCR site-directed, mutagenesis. Comparison of the kinetic parameters of these mutants with, those of the wild-type aspartate kinase suggested that Thr47 was involved, in binding aspartate and that Lys7 and Glu74 were involved in catalysis., Analysis of the effective concentrations of magnesium ion on the activity, showed that the mutants with replacements at Ser41, Thr47, Asp154 and, Asp182 required higher concentrations of magnesium ion. This suggests that, these four residues play important roles in the binding of magnesium ions, which are required for enzymatic activity.
+To reveal the catalytic mechanism of Thermus aspartate kinase, each of 29 amino acid residues that were highly conserved in the sequenced aspartate kinases, was replaced with alanine or leucine by PCR site-directed mutagenesis. Comparison of the kinetic parameters of these mutants with those of the wild-type aspartate kinase suggested that Thr47 was involved in binding aspartate and that Lys7 and Glu74 were involved in catalysis. Analysis of the effective concentrations of magnesium ion on the activity showed that the mutants with replacements at Ser41, Thr47, Asp154 and Asp182 required higher concentrations of magnesium ion. This suggests that these four residues play important roles in the binding of magnesium ions which are required for enzymatic activity.
 ==About this Structure==
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 [[Category: regulatory subunit]]
-''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Wed Jan 23 14:30:00 2008''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 17:02:28 2008''