The crystal structure of fructose‐1,6‐bisphosphate aldolase fromDrosophila melanogaster at 2.5A˚resolution

Abstract

The structure of fructose-l,6-bisphosphate aldolase from Drosophila melanogaster has been determined by X-my diffraction at 2.5 .~ resolution.The insect enzyme crystallizes in space group P'2r2~2t with lattice constants a=86.60, b= 116.80, c= 151.58 ~. Molecular replacement with rabbit muscle aldolase as a search model has been employed to solve the structure. To improve the initial phases real space averaging, including phase extension from 4.0 to 2.5 ,/~, has been applied. Refinement of the atomic positions by molecular dynamics resulted in a crystallographic R-factor of 0.214. The tertiary structure resembles in most parts that of the vertebrate aldolase from rabbit muscle. Significant differences were found in surface loops and the N-and C-terminal regions of the protein. Here we present the first aldolase structure where the functionally important C-terminal arm is described completely.Crystal structure; Glycolytic enzyme; Fructose-1,6-bisphosphate aldolase; Substrate specificity; Drosophila melanogaster INTRC, DUCTIONFruetose-l,6-bisphosphate (FBP) aldolase (EC 4.1.2.13) is a glycolytic enzyme catalyzing the reversible aldol cleavage of fructose-l,6-bisphosphate to dihydroxyacetone phosphate and D-glyceraldehyde-3-phosphate. FBP aldolase from Drosophila melanogaster (strain Sevelen, wild type, pupea) forms a Schiff base between the substrate and a lysyl residue [1] and belongs therefore to the class I aldolases [2]. Contrary to class I the class II aldolases use a divalent metal ion as cofactor [3]. The vertebrate aldolases (class I) comprise three isozymes. They refer to A (muscle), B (liver) and C (brain) depending on the tissue from which they are isolated [3]. The A and C form show a higher substrate specificity for FBP over fructose-l-phosphate (F1P), while the liver isozyme has no such preference.Abbreviations: DALD, fructose-l,6-bisphosphate aldolase from Drosophila melanogaster; RMALD, rabbit muscle aldolase; HMALD, human muscle aldolase; RLALD, rabbit liver aldolase; FBP, fructose-1,6-bisphosphate; FIP, fructose-l-phosphate; r.m.s., root-meansquare.Sequence numbering: Throughout this paper the sequence numbering of FBP aldolase from rabbit muscle will be applied.

A Simple Cross-Linking/Mass Spectrometry Workflow for Studying System-wide Protein Interactions

“…To exemplify this often-overlooked limitation of the XLMS technique, we tentatively mapped cross-links in the structural model of aldolase (PDB entry 2pyo). Drosophila aldolase is a homotetramer of 158 kDa catalyzing the aldol cleavage of fructose l,6-bisphosphate . We identified a total of 10 cross-links in aldolase of Drosophila embryos.…”

Dynamic Enolate Recognition in Aqueous Solution by Zinc(II) in a Phenacyl-Pendant Cyclen Complex: Implications for the Role of Zinc(II) in Class II Aldolases

“…Aldolases are classified into two groups on the basis of reaction mechanism. Class I aldolases, which occur in animals and plants, form a Schiff base intermediate between the ε-amino group of the lysine residue and the carbonyl group of the sugar .…”

Mechanism of the Schiff Base Forming Fructose-1,6-bisphosphate Aldolase: Structural Analysis of Reaction Intermediates

“…FBPA I is grouped into two different sequence families, one with only eukaryotic members and one with both bacterial and archaeal members (termed the archaeal FBPA I or FBPA IA) (). Several crystal structures have shown that the eukaryotic FBPA I is a homotetramer, where the subunits adopt the fold of a parallel (βα) 8 -(TIM)-barrel ( − ). Although the archaeal FBPA I subunit also displays the (βα) 8 -barrel fold, its quaternary structure is that of a homodecamer resulting from the dimerization of two identical pentamers ().…”