Human aldolase A natural mutants: relationship between flexibility of the C-terminal region and enzyme function (604 views)
Biochem J (ISSN: 0264-6021, 1470-8728electronic, 0264-6021linking), 2004 May 15; 380: 51-56.
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Paper type: Journal Article,
Impact factor: 4.278, 5-year impact factor: 4.95
Url: http://www.scopus.com/inward/record.url?eid=2-s2.0-2642534395&partnerID=40&md5=9a67fef59be66b14431663cb5d63cbe7
Keywords: Aldolase A, Aldolase A Gene Mutation, Aldolase A Mutant Expression, Fructose 1, 6-Bisphosphate, Molecular Modelling, Bacteria, Conformations, Enzymes, Molecular Graphics, Anemia, Myopathic Symptoms, Biochemistry, Complementary Dna, 6 Bisphosphate, Fructose Bisphosphate Aldolase, Hydrolase, Mutant Protein, Arthrogryposis, Article, Birth, Carboxy Terminal Sequence, Controlled Study, Crystallography, Ectopic Tissue, Enzyme Activity, Enzyme Deficiency, Hemolytic Anemia, Heterozygosity, Human, Melting Point, Priority Journal, Protein Analysis, Protein Expression, Protein Function, School Child, Sequence Analysis, Substitution Reaction, Temperature Measurement, Wild Type, Amino Acid Substitution, Congenital, Circular Dichroism, Codon, Fructose-Bisphosphate Aldolase, Glycine, Infant, Kinetics, Molecular Weight, Muscle Weakness, Mutagenesis, Site-Directed, Missense, Point Mutation, Protein Conformation, Protein Denaturation, Protein Structure, Tertiary, Recombinant Fusion Proteins, Structure-Activity Relationship, Bacteria (microorganisms), Felis Catus, Oryctolagus Cuniculus,
Affiliations:
*** IBB - CNR ***
Dipto. Biochim. Biotecnologie M., Univ. di Napoli Federico II, Via S. Pansini 5, I-80131 Napoli, Italy
CEINGE-Biotecnologie Avanzate, Univ. di Napoli Federico II, Via S. Pansini 5, I-80131 Napoli, Italy
Ist. di Biostrutture e Bioimmagini, CNR, Via Mezzocannone 6, I-80134 Napoli, Italy
Dipartimento di Pediatria, Policlinico, Università di Catania, Via S. Sofia 78, I-95123 Catania, Italy
Dipartimento di Chimica Biologica, Univ. di Napoli Federico II, Via Mezzocannone 6, I-80134 Napoli, Italy
References:
Not available.
Biochem J (ISSN: 0264-6021, 1470-8728electronic, 0264-6021linking), 2004 May 15; 380: 51-56.
Export to BibTeX Export to EndNote
Paper type: Journal Article,
Impact factor: 4.278, 5-year impact factor: 4.95
Url: http://www.scopus.com/inward/record.url?eid=2-s2.0-2642534395&partnerID=40&md5=9a67fef59be66b14431663cb5d63cbe7
Keywords: Aldolase A, Aldolase A Gene Mutation, Aldolase A Mutant Expression, Fructose 1, 6-Bisphosphate, Molecular Modelling, Bacteria, Conformations, Enzymes, Molecular Graphics, Anemia, Myopathic Symptoms, Biochemistry, Complementary Dna, 6 Bisphosphate, Fructose Bisphosphate Aldolase, Hydrolase, Mutant Protein, Arthrogryposis, Article, Birth, Carboxy Terminal Sequence, Controlled Study, Crystallography, Ectopic Tissue, Enzyme Activity, Enzyme Deficiency, Hemolytic Anemia, Heterozygosity, Human, Melting Point, Priority Journal, Protein Analysis, Protein Expression, Protein Function, School Child, Sequence Analysis, Substitution Reaction, Temperature Measurement, Wild Type, Amino Acid Substitution, Congenital, Circular Dichroism, Codon, Fructose-Bisphosphate Aldolase, Glycine, Infant, Kinetics, Molecular Weight, Muscle Weakness, Mutagenesis, Site-Directed, Missense, Point Mutation, Protein Conformation, Protein Denaturation, Protein Structure, Tertiary, Recombinant Fusion Proteins, Structure-Activity Relationship, Bacteria (microorganisms), Felis Catus, Oryctolagus Cuniculus,
Affiliations:
*** IBB - CNR ***
Dipto. Biochim. Biotecnologie M., Univ. di Napoli Federico II, Via S. Pansini 5, I-80131 Napoli, Italy
CEINGE-Biotecnologie Avanzate, Univ. di Napoli Federico II, Via S. Pansini 5, I-80131 Napoli, Italy
Ist. di Biostrutture e Bioimmagini, CNR, Via Mezzocannone 6, I-80134 Napoli, Italy
Dipartimento di Pediatria, Policlinico, Università di Catania, Via S. Sofia 78, I-95123 Catania, Italy
Dipartimento di Chimica Biologica, Univ. di Napoli Federico II, Via Mezzocannone 6, I-80134 Napoli, Italy
References:
Not available.
Human aldolase A natural mutants: relationship between flexibility of the C-terminal region and enzyme function
We have identified a new mutation in the FBP (fructose 1,6-bis-phosphate) aldolase A gene in a child with suspected haemolytic anaemia associated with myopathic symptoms at birth and with a subsequent diagnosis of arthrogryposis multiplex congenita and pituitary ectopia. Sequence analysis of the whole gene, also performed on the patient's full-length cDNA, revealed only a Gly(346) --> Ser substitution in the heterozygous state. We expressed in a bacterial system the new aldolase A Gly(346) --> Ser mutant, and the Glu(206) --> Lys mutant identified by others, in a patient with an aldolase A deficit. Analysis of their functional profiles showed that the Gly(346) --> Ser mutant had the same K-m as the wild-type enzyme, but a 4-fold lower k(cat). The Glu(206) --> Lys mutant had a K-m approx. 2-fold higher than that of both the Gly(346) --> Ser mutant and the wild-type enzyme, and a k(cat) value 40 % less than the wild-type. The Gly(346) --> Ser and wild-type enzymes had the same T-m (melting temperature), which was approx. 6-7 degreesC higher than that of the Glu(206) --> Lys enzyme. An extensive molecular graphic analysis of the mutated enzymes, using human and rabbit aldolase A crystallographic structures, suggests that the Glu(206) --> Lys mutation destabilizes the aldolase A tetramer at the subunit interface, and highlights the fact that the glycine-to-serine substitution at position 346 limits the flexibility of the C-terminal region. These results also provide the first evidence that Gly(346) is crucial for the correct conformation and function of aldolase A, because it governs the entry/release of the substrates into/from the enzyme cleft, and/or allows important C-terminal residues to approach the active site.
We have identified a new mutation in the FBP (fructose 1,6-bis-phosphate) aldolase A gene in a child with suspected haemolytic anaemia associated with myopathic symptoms at birth and with a subsequent diagnosis of arthrogryposis multiplex congenita and pituitary ectopia. Sequence analysis of the whole gene, also performed on the patient's full-length cDNA, revealed only a Gly(346) --> Ser substitution in the heterozygous state. We expressed in a bacterial system the new aldolase A Gly(346) --> Ser mutant, and the Glu(206) --> Lys mutant identified by others, in a patient with an aldolase A deficit. Analysis of their functional profiles showed that the Gly(346) --> Ser mutant had the same K-m as the wild-type enzyme, but a 4-fold lower k(cat). The Glu(206) --> Lys mutant had a K-m approx. 2-fold higher than that of both the Gly(346) --> Ser mutant and the wild-type enzyme, and a k(cat) value 40 % less than the wild-type. The Gly(346) --> Ser and wild-type enzymes had the same T-m (melting temperature), which was approx. 6-7 degreesC higher than that of the Glu(206) --> Lys enzyme. An extensive molecular graphic analysis of the mutated enzymes, using human and rabbit aldolase A crystallographic structures, suggests that the Glu(206) --> Lys mutation destabilizes the aldolase A tetramer at the subunit interface, and highlights the fact that the glycine-to-serine substitution at position 346 limits the flexibility of the C-terminal region. These results also provide the first evidence that Gly(346) is crucial for the correct conformation and function of aldolase A, because it governs the entry/release of the substrates into/from the enzyme cleft, and/or allows important C-terminal residues to approach the active site.
Human aldolase A natural mutants: relationship between flexibility of the C-terminal region and enzyme function
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Human aldolase A natural mutants: relationship between flexibility of the C-terminal region and enzyme function
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Andreotti G, Pedone E, Giordano A, Cubellis MV
* Biochemical phenotype of a common disease-causing mutation and a possible therapeutic approach for the phosphomannomutase 2-associated disorder of glycosylation (515 views)
Molecular Genetics & Genomic Medicine (ISSN: 2324-9269), 2013 May; 1(1): 32-44.
Impact Factor: 2.082
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Esposito G, Vitagliano L, Cevenini A, Amelio T, Zagari A, Salvatore F
* Unraveling the structural and functional features of an aldolase A mutant involved in the hemolytic anemia and severe rhabdomyolysis reported in a child (984 views)
Haematologica (ISSN: 0006-4971, 0006-6497, 0390-6078), 2005 Jan 15; 105(2): 905-906.
Impact Factor: 10.131
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* Biochemical phenotype of a common disease-causing mutation and a possible therapeutic approach for the phosphomannomutase 2-associated disorder of glycosylation (515 views)
Molecular Genetics & Genomic Medicine (ISSN: 2324-9269), 2013 May; 1(1): 32-44.
Impact Factor: 2.082
View Export to BibTeX Export to EndNote
Esposito G, Vitagliano L, Cevenini A, Amelio T, Zagari A, Salvatore F
* Unraveling the structural and functional features of an aldolase A mutant involved in the hemolytic anemia and severe rhabdomyolysis reported in a child (984 views)
Haematologica (ISSN: 0006-4971, 0006-6497, 0390-6078), 2005 Jan 15; 105(2): 905-906.
Impact Factor: 10.131
View Export to BibTeX Export to EndNote
2 Records (2 excluding Abstracts).
Total impact factor: 12.213 (12.213 excluding Abstracts).
Total 5 year impact factor: 11.855 (11.855 excluding Abstracts).
Total impact factor: 12.213 (12.213 excluding Abstracts).
Total 5 year impact factor: 11.855 (11.855 excluding Abstracts).
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