Dual Coenzyme Specificity of Photosynthetic Glyceraldehyde-3-phosphate Dehydrogenase Interpreted by the Crystal Structure of A4 Isoform Complexed with NAD
| Autor: | Francesca Sparla, Giuseppe Falini, Piera Sabatino, Simona Fermani, Paolo Trost, Alberto Ripamonti, Paolo Pupillo |
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| Rok vydání: | 2003 |
| Předmět: |
Models
Molecular Macromolecular Substances Stereochemistry Molecular Sequence Data Dehydrogenase Crystallography X-Ray Photosynthesis Biochemistry Cofactor chemistry.chemical_compound stomatognathic system Spinacia oleracea Oxidoreductase Ribose Amino Acid Sequence Glyceraldehyde 3-phosphate dehydrogenase chemistry.chemical_classification Binding Sites biology NAD Recombinant Proteins Amino acid Isoenzymes Kinetics chemistry biology.protein NAD+ kinase Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) Sequence Alignment NADP |
| Zdroj: | Biochemistry. 42:4631-4639 |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/bi0272149 |
| Popis: | Photosynthetic glyceraldehyde-3-phosphate dehydrogenase (GAPDH) of Spinacia oleracea belongs to a wide group of GAPDHs found in most organisms displaying oxygenic photosynthesis, including cyanobacteria, green and red algae, and higher plants. As a major catalytic difference with respect to glycolytic GAPDH, photosynthetic GAPDH exhibits dual cofactor specificity toward pyridine nucleotides with a preference for NADP(H). Here we report the crystal structure of NAD-complexed recombinant A(4)-GAPDH (NAD-A(4)-GAPDH) from Spinacia oleracea, expressed in Escherichia coli. Its superimposition onto native A(4)-GAPDH complexed with NADP (NADP-A(4)-GAPDH) pinpoints specific conformational changes resulting from cofactor replacement. In photosynthetic NAD-A(4)-GAPDH, the side chain of Asp32 is oriented toward the coenzyme to interact with the adenine ribose diol, similar to glycolytic GAPDHs (NAD-specific). On the contrary, in NADP-A(4)-GAPDH Asp32 moves away to accommodate the additional 2'-phosphate group of the coenzyme and to minimize electrostatic repulsion. Asp32 rotation is allowed by the presence of the small residue Ala40, conserved in most photosynthetic GAPDHs, replacing bulky amino acid side chains in glycolytic GAPDHs. While in NADP-A(4)-GAPDH two amino acids, Thr33 and Ser188, are involved in hydrogen bonds with the 2'-phosphate group of NADP, in the NAD-complexed enzyme these interactions are lacking. The crystallographic structure of NAD-A(4)-GAPDH highlights that four residues, Thr33, Ala40, Ser188, and Ala187 (Leu, Leu, Pro, and Leu respectively, in glycolytic Bacillus stearothermophilus GAPDH sequence) are of primary importance for the dual cofactor specificity of photosynthetic GAPDH. These modifications seem to trace the minimum evolutionary route for a primitive NAD-specific GAPDH to be converted into the NADP-preferring enzyme of oxygenic photosynthetic organisms. |
| Databáze: | OpenAIRE |
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