Carboxy-terminus Recruitment Induced by Substrate Binding in Eukaryotic Fructose Bis-phosphate Aldolases.
From: Département de Biochimie, Université de Montréal, Montréal, Québec H3C 3J7, Canada.
Biochemistry
- Publish Date: Aug 2007
- ISSN: 0006-2960
- Volume: 46
- Issue: 33
- Pages: 9533-40
- Medium: Print
- Language: English
- Citation (JAMA): Lafrance-Vanasse Julien, Sygusch Jurgen, et al. Carboxy-terminus Recruitment Induced by Substrate Binding in Eukaryotic Fructose Bis-phosphate Aldolases.. Biochemistry Aug 2007;46:9533-40
Abstract
The crystal structures of Leishmania mexicana fructose-1,6-bis(phosphate) aldolase in complex with substrate and competitive inhibitor, mannitol-1,6-bis(phosphate), were solved to 2.2 A resolution. Crystallographic analysis revealed a Schiff base intermediate trapped in the native structure complexed with substrate while the inhibitor was trapped in a conformation mimicking the carbinolamine intermediate. Binding modes corroborated previous structures reported for rabbit muscle aldolase. Amino acid substitution of Gly-312 to Ala, adjacent to the P1-phosphate binding site and unique to trypanosomatids, did not perturb ligand binding in the active site. Ligand attachment ordered amino acid residues 359-367 of the C-terminal region (353-373) that was disordered beyond Asp-358 in the unbound structure, revealing a novel recruitment mechanism of this region by aldolases. C-Terminal peptide ordering is triggered by P1-phosphate binding that induces conformational changes whereby C-terminal Leu-364 contacts P1-phosphate binding residue Arg-313. C-Terminal region capture synergizes additional interactions with subunit surface residues, not perturbed by P1-phosphate binding, and stabilizes C-terminal attachment. Amino acid residues that participate in the capturing interaction are conserved among class I aldolases, indicating a general recruitment mechanism whereby C-terminal capture facilitates active site interactions in subsequent catalytic steps. Recruitment accelerates the enzymatic reaction by using binding energy to reduce configurational entropy during catalysis thereby localizing the conserved C-terminus tyrosine, which mediates proton transfer, proximal to the active site enamine.
Mesh Headings (Keywords): Alanine, Amino Acid Sequence, Amino Acid Substitution, Animals, Binding Sites, Crystallography, X-Ray, Enzyme Inhibitors, Fructose-Bisphosphate Aldolase, Glycine, Leishmania mexicana, Mannitol Phosphates, Molecular Sequence Data, Protozoan Proteins, Rabbits, Substrate Specificity
Check for Full Text / PubMed Unique Identifier (PMID): 17661446
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