2vgf: Difference between revisions
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==Overview== | ==Overview== | ||
Deficiency of human erythrocyte isozyme (RPK) is, together with | Deficiency of human erythrocyte isozyme (RPK) is, together with glucose-6-phosphate dehydrogenase deficiency, the most common cause of the nonspherocytic hemolytic anemia. To provide a molecular framework to the disease, we have solved the 2.7 A resolution crystal structure of human RPK in complex with fructose 1,6-bisphosphate, the allosteric activator, and phosphoglycolate, a substrate analogue, and we have functionally and structurally characterized eight mutants (G332S, G364D, T384M, D390N, R479H, R486W, R504L, and R532W) found in RPK-deficient patients. The mutations target distinct regions of RPK structure, including domain interfaces and catalytic and allosteric sites. The mutations affect to a different extent thermostability, catalytic efficiency, and regulatory properties. These studies are the first to correlate the clinical symptoms with the molecular properties of the mutant enzymes. Mutations greatly impairing thermostability and/or activity are associated with severe anemia. Some mutant proteins exhibit moderate changes in the kinetic parameters, which are sufficient to cause mild to severe anemia, underlining the crucial role of RPK for erythrocyte metabolism. Prediction of the effects of mutations is difficult because there is no relation between the nature and location of the replaced amino acid and the type of molecular perturbation. Characterization of mutant proteins may serve as a valuable tool to assist with diagnosis and genetic counseling. | ||
==About this Structure== | ==About this Structure== | ||
2VGF is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with <scene name='pdbligand=FBP:'>FBP</scene>, <scene name='pdbligand=PGA:'>PGA</scene>, <scene name='pdbligand=K:'>K</scene> and <scene name='pdbligand=MN:'>MN</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. This structure | 2VGF is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with <scene name='pdbligand=FBP:'>FBP</scene>, <scene name='pdbligand=PGA:'>PGA</scene>, <scene name='pdbligand=K:'>K</scene> and <scene name='pdbligand=MN:'>MN</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. This structure supersedes the now removed PDB entry 1LIW. Active as [http://en.wikipedia.org/wiki/Pyruvate_kinase Pyruvate kinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.1.40 2.7.1.40] Known structural/functional Sites: <scene name='pdbsite=AC1:Fbp+Binding+Site+For+Chain+A'>AC1</scene>, <scene name='pdbsite=AC2:Pga+Binding+Site+For+Chain+A'>AC2</scene>, <scene name='pdbsite=AC3:K+Binding+Site+For+Chain+A'>AC3</scene>, <scene name='pdbsite=AC4:Mn+Binding+Site+For+Chain+A'>AC4</scene>, <scene name='pdbsite=AC5:Fbp+Binding+Site+For+Chain+B'>AC5</scene>, <scene name='pdbsite=AC6:Pga+Binding+Site+For+Chain+B'>AC6</scene>, <scene name='pdbsite=AC7:K+Binding+Site+For+Chain+B'>AC7</scene>, <scene name='pdbsite=AC8:Mn+Binding+Site+For+Chain+B'>AC8</scene>, <scene name='pdbsite=AC9:Fbp+Binding+Site+For+Chain+C'>AC9</scene>, <scene name='pdbsite=BC1:Pga+Binding+Site+For+Chain+C'>BC1</scene>, <scene name='pdbsite=BC2:K+Binding+Site+For+Chain+C'>BC2</scene>, <scene name='pdbsite=BC3:Mn+Binding+Site+For+Chain+C'>BC3</scene>, <scene name='pdbsite=BC4:Fbp+Binding+Site+For+Chain+D'>BC4</scene>, <scene name='pdbsite=BC5:Pga+Binding+Site+For+Chain+D'>BC5</scene>, <scene name='pdbsite=BC6:K+Binding+Site+For+Chain+D'>BC6</scene> and <scene name='pdbsite=BC7:Mn+Binding+Site+For+Chain+D'>BC7</scene>. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2VGF OCA]. | ||
==Reference== | ==Reference== | ||
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[[Category: Pyruvate kinase]] | [[Category: Pyruvate kinase]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Abraham, D | [[Category: Abraham, D J.]] | ||
[[Category: Bianchi, P.]] | [[Category: Bianchi, P.]] | ||
[[Category: Chiarelli, L.]] | [[Category: Chiarelli, L.]] | ||
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[[Category: transferase]] | [[Category: transferase]] | ||
''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 18:55:42 2008'' | ||
Revision as of 19:55, 21 February 2008
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HUMAN ERYTHROCYTE PYRUVATE KINASE: T384M MUTANT
OverviewOverview
Deficiency of human erythrocyte isozyme (RPK) is, together with glucose-6-phosphate dehydrogenase deficiency, the most common cause of the nonspherocytic hemolytic anemia. To provide a molecular framework to the disease, we have solved the 2.7 A resolution crystal structure of human RPK in complex with fructose 1,6-bisphosphate, the allosteric activator, and phosphoglycolate, a substrate analogue, and we have functionally and structurally characterized eight mutants (G332S, G364D, T384M, D390N, R479H, R486W, R504L, and R532W) found in RPK-deficient patients. The mutations target distinct regions of RPK structure, including domain interfaces and catalytic and allosteric sites. The mutations affect to a different extent thermostability, catalytic efficiency, and regulatory properties. These studies are the first to correlate the clinical symptoms with the molecular properties of the mutant enzymes. Mutations greatly impairing thermostability and/or activity are associated with severe anemia. Some mutant proteins exhibit moderate changes in the kinetic parameters, which are sufficient to cause mild to severe anemia, underlining the crucial role of RPK for erythrocyte metabolism. Prediction of the effects of mutations is difficult because there is no relation between the nature and location of the replaced amino acid and the type of molecular perturbation. Characterization of mutant proteins may serve as a valuable tool to assist with diagnosis and genetic counseling.
About this StructureAbout this Structure
2VGF is a Single protein structure of sequence from Homo sapiens with , , and as ligands. This structure supersedes the now removed PDB entry 1LIW. Active as Pyruvate kinase, with EC number 2.7.1.40 Known structural/functional Sites: , , , , , , , , , , , , , , and . Full crystallographic information is available from OCA.
ReferenceReference
Structure and function of human erythrocyte pyruvate kinase. Molecular basis of nonspherocytic hemolytic anemia., Valentini G, Chiarelli LR, Fortin R, Dolzan M, Galizzi A, Abraham DJ, Wang C, Bianchi P, Zanella A, Mattevi A, J Biol Chem. 2002 Jun 28;277(26):23807-14. Epub 2002 Apr 17. PMID:11960989
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Proteopedia Page Contributors and Editors (what is this?)Proteopedia Page Contributors and Editors (what is this?)
OCA- Pages with broken file links
- Homo sapiens
- Pyruvate kinase
- Single protein
- Abraham, D J.
- Bianchi, P.
- Chiarelli, L.
- Dolzan, M.
- Fortin, R.
- Galizzi, A.
- Mattevi, A.
- Valentini, G.
- Wang, C.
- Zanella, A.
- FBP
- K
- MN
- PGA
- Alternative splicing
- Disease mutation
- Glycolysis
- Kinase
- Magnesium
- Metal-binding
- Phosphorylation
- Polymorphism
- Pyruvate
- Pyruvate kinase in the active r-state
- Transferase