1w7w
From Proteopedia
(New page: 200px<br /><applet load="1w7w" size="450" color="white" frame="true" align="right" spinBox="true" caption="1w7w, resolution 2.80Å" /> '''STRUCTURE AND MUTATI...) |
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| - | [[Image:1w7w.gif|left|200px]]<br /><applet load="1w7w" size=" | + | [[Image:1w7w.gif|left|200px]]<br /><applet load="1w7w" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1w7w, resolution 2.80Å" /> | caption="1w7w, resolution 2.80Å" /> | ||
'''STRUCTURE AND MUTATIONAL ANALYSIS OF A PLANT MITOCHONDRIAL NUCLEOSIDE DIPHOSPHATE KINASE: IDENTIFICATION OF RESIDUES INVOLVED IN SERINE PHOSPHORYLATION AND OLIGOMERIZATION.'''<br /> | '''STRUCTURE AND MUTATIONAL ANALYSIS OF A PLANT MITOCHONDRIAL NUCLEOSIDE DIPHOSPHATE KINASE: IDENTIFICATION OF RESIDUES INVOLVED IN SERINE PHOSPHORYLATION AND OLIGOMERIZATION.'''<br /> | ||
==Overview== | ==Overview== | ||
| - | We report the first crystal structure of a plant (Pisum sativum L. cv | + | We report the first crystal structure of a plant (Pisum sativum L. cv Oregon sugarpod) mitochondrial nucleoside diphosphate kinase. Similar to other eukaryotic nucleoside diphosphate kinases, the plant enzyme is a hexamer; the six monomers in the asymmetric unit are arranged as trimers of dimers. Different functions of the kinase have been correlated with the oligomeric structure and the phosphorylation of Ser residues. We show that the occurrence of Ser autophosphorylation depends on enzymatic activity. The mutation of the strictly conserved Ser-119 to Ala reduced the Ser phosphorylation to about one-half of that observed in wild type with only a modest change of enzyme activity. We also show that mutating another strictly conserved Ser, Ser-69, to Ala reduces the enzyme activity to 6% and 14% of wild-type using dCDP and dTDP as acceptors, respectively. Changes in the oligomerization pattern of the S69A mutant were observed by cross-linking experiments. A reduction in trimer formation and a change in the dimer interaction could be detected with a concomitant increase of tetramers. We conclude that the S69 mutant is involved in the stabilization of the oligomeric state of this plant nucleoside diphosphate kinase. |
==About this Structure== | ==About this Structure== | ||
| - | 1W7W is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Pisum_sativum Pisum sativum]. Active as [http://en.wikipedia.org/wiki/Nucleoside-diphosphate_kinase Nucleoside-diphosphate kinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.4.6 2.7.4.6] Full crystallographic information is available from [http:// | + | 1W7W is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Pisum_sativum Pisum sativum]. Active as [http://en.wikipedia.org/wiki/Nucleoside-diphosphate_kinase Nucleoside-diphosphate kinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.4.6 2.7.4.6] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1W7W OCA]. |
==Reference== | ==Reference== | ||
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[[Category: transferase]] | [[Category: transferase]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 15:41:26 2008'' |
Revision as of 13:41, 21 February 2008
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STRUCTURE AND MUTATIONAL ANALYSIS OF A PLANT MITOCHONDRIAL NUCLEOSIDE DIPHOSPHATE KINASE: IDENTIFICATION OF RESIDUES INVOLVED IN SERINE PHOSPHORYLATION AND OLIGOMERIZATION.
Overview
We report the first crystal structure of a plant (Pisum sativum L. cv Oregon sugarpod) mitochondrial nucleoside diphosphate kinase. Similar to other eukaryotic nucleoside diphosphate kinases, the plant enzyme is a hexamer; the six monomers in the asymmetric unit are arranged as trimers of dimers. Different functions of the kinase have been correlated with the oligomeric structure and the phosphorylation of Ser residues. We show that the occurrence of Ser autophosphorylation depends on enzymatic activity. The mutation of the strictly conserved Ser-119 to Ala reduced the Ser phosphorylation to about one-half of that observed in wild type with only a modest change of enzyme activity. We also show that mutating another strictly conserved Ser, Ser-69, to Ala reduces the enzyme activity to 6% and 14% of wild-type using dCDP and dTDP as acceptors, respectively. Changes in the oligomerization pattern of the S69A mutant were observed by cross-linking experiments. A reduction in trimer formation and a change in the dimer interaction could be detected with a concomitant increase of tetramers. We conclude that the S69 mutant is involved in the stabilization of the oligomeric state of this plant nucleoside diphosphate kinase.
About this Structure
1W7W is a Single protein structure of sequence from Pisum sativum. Active as Nucleoside-diphosphate kinase, with EC number 2.7.4.6 Full crystallographic information is available from OCA.
Reference
Structure and mutational analysis of a plant mitochondrial nucleoside diphosphate kinase. Identification of residues involved in serine phosphorylation and oligomerization., Johansson M, Mackenzie-Hose A, Andersson I, Knorpp C, Plant Physiol. 2004 Oct;136(2):3034-42. Epub 2004 Oct 1. PMID:15466238
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