A gatekeeper helix determines the substrate specificity of Sjögren–Larsson Syndrome enzyme fatty aldehyde dehydrogenase

Keller, Markus A.; Zander, U.; Fuchs, Julian E.; Kreutz, Christoph; Watschinger, Katrin; Mueller, Thomas; Golderer, Georg; Liedl, Klaus R.; Ralser, Markus; Kräutler, Bernhard; Werner, Ernst R.; Márquez, José Antonio

doi:10.1038/ncomms5439

Cited by 58 publications

(55 citation statements)

References 70 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…An even longer tail at the С-terminus was observed only in human fatty aldehyde dehydrogenase (PDB entry 4QGK, [38]). In the latter structure, the α -helix at the C-terminus was shown to cover the substrate channel entrance, acting as a gatekeeper.…”

Section: Resultsmentioning

confidence: 99%

NADP-Dependent Aldehyde Dehydrogenase from ArchaeonPyrobaculum sp.1860: Structural and Functional Features

Bezsudnova

Petrova

Artemova

et al. 2016

Archaea

View full text Add to dashboard Cite

We present the functional and structural characterization of the first archaeal thermostable NADP-dependent aldehyde dehydrogenase AlDHPyr1147. In vitro, AlDHPyr1147 catalyzes the irreversible oxidation of short aliphatic aldehydes at 60–85°С, and the affinity of AlDHPyr1147 to the NADP+ at 60°С is comparable to that for mesophilic analogues at 25°С. We determined the structures of the apo form of AlDHPyr1147 (3.04 Å resolution), three binary complexes with the coenzyme (1.90, 2.06, and 2.19 Å), and the ternary complex with the coenzyme and isobutyraldehyde as a substrate (2.66 Å). The nicotinamide moiety of the coenzyme is disordered in two binary complexes, while it is ordered in the ternary complex, as well as in the binary complex obtained after additional soaking with the substrate. AlDHPyr1147 structures demonstrate the strengthening of the dimeric contact (as compared with the analogues) and the concerted conformational flexibility of catalytic Cys287 and Glu253, as well as Leu254 and the nicotinamide moiety of the coenzyme. A comparison of the active sites of AlDHPyr1147 and dehydrogenases characterized earlier suggests that proton relay systems, which were previously proposed for dehydrogenases of this family, are blocked in AlDHPyr1147, and the proton release in the latter can occur through the substrate channel.

show abstract

Section: Resultsmentioning

confidence: 99%

NADP-Dependent Aldehyde Dehydrogenase from ArchaeonPyrobaculum sp.1860: Structural and Functional Features

Bezsudnova

Petrova

Artemova

et al. 2016

Archaea

View full text Add to dashboard Cite

show abstract

“…Several endogenous enzymatic detoxification enzymes, such as glutathione-s-transferase and aldehyde dehydrogenase, are present in tissues to efficiently neutralize the insidious effects of lipid aldehydes. It has been noted that ( 2E )-hexadecenal is principally detoxified by microsomal NAD+ dependent fatty aldehyde dehydrogenase (FALDH) enzyme (Keller et al, 2014). Genetic mutations of FALDH cause Sjogren-Larsson Syndrom (SLS), a rare epithelial and neurological disorder.…”

Section: Resultsmentioning

confidence: 99%

A facile stable-isotope dilution method for determination of sphingosine phosphate lyase activity

Suh

Eltanawy

Rangan

et al. 2016

Chemistry and Physics of Lipids

View full text Add to dashboard Cite

A new technique for quantifying sphingosine phosphate lyase activity in biological samples is described. In this procedure, 2-hydrazinoquinoline is used to convert (2E)-hexadecenal into the corresponding hydrazone derivative to improve ionization efficiency and selectivity of detection. Combined utilization of liquid chromatographic separation and multiple reaction monitoring-mass spectrometry allows for simultaneous quantification of the substrate S1P and product (2E)-hexadecenal. Incorporation of (2E)-d5-hexadecenal as an internal standard improves detection accuracy and precision. A simple one-step derivatization procedure eliminates the need for further extractions. Limits of quantification for (2E)-hexadecenal and sphingosine-1-phosphate are 100 and 50 fmol, respectively. The assay displays a wide dynamic detection range useful for detection of low basal sphingosine phosphate lyase activity in wild type cells, SPL-overexpressing cell lines, and wild type mouse tissues. Compared to current methods, the capacity for simultaneous detection of sphingosine-1-phosphate and (2E)-hexadecenal greatly improves the accuracy of results and shows excellent sensitivity and specificity for sphingosine phosphate lyase activity detection.

show abstract

“…During our work, a detailed X-ray structure and mechanistic study of the human FALDH was published with a view to understanding the substrate specificity of the enzyme and the impact of mutations that cause Sjogren-Larsson syndrome (54). In this report, the FALDH construct contained an N-terminal streptavidin affinity tag and, in comparison to our truncation, only removed the C-terminal 22 amino acids, the minimum based on the proposed TM region.…”

Section: Discussionmentioning

confidence: 99%

Characterization of homologous sphingosine-1-phosphate lyase isoforms in the bacterial pathogen Burkholderia pseudomallei

McLean

Marles‐Wright

Custódio

et al. 2017

Journal of Lipid Research

View full text Add to dashboard Cite

Sphingolipids (SLs) are ubiquitous elements in eukaryotic membranes and are also found in some bacterial and viral species. As well as playing an integral structural role, SLs also act as potent signaling molecules involved in numerous cellular pathways and have been linked to many human diseases. A central SL signaling molecule is sphingosine-1-phosphate (S1P), whose breakdown is catalyzed by S1P lyase (S1PL), a pyridoxal 5′-phosphate (PLP)-dependent enzyme that catalyzes the cleavage of S1P to (2E)-hexadecenal (2E-HEX) and phosphoethanolamine. Here, we show that the pathogenic bacterium, Burkholderia pseudomallei K96243, encodes two homologous proteins (S1PL2021 and S1PL2025) that display moderate sequence identity to known eukaryotic and prokaryotic S1PLs. Using an established MS-based methodology, we show that recombinant S1PL2021 is catalytically active. We also used recombinant human fatty aldehyde dehydrogenase to develop a spectrophotometric enzyme-coupled assay to detect 2E-HEX formation and measure the kinetic constants of the two B. pseudomallei S1PL isoforms. Furthermore, we determined the X-ray crystal structure of the PLP-bound form of S1PL2021 at 2.1 Å resolution revealing that the enzyme displays a conserved structural fold and active site architecture comparable with known S1PLs. The combined data suggest that B. pseudomallei has the potential to degrade host SLs in a S1PL-dependent manner.

show abstract

A gatekeeper helix determines the substrate specificity of Sjögren–Larsson Syndrome enzyme fatty aldehyde dehydrogenase

Cited by 58 publications

References 70 publications

NADP-Dependent Aldehyde Dehydrogenase from ArchaeonPyrobaculum sp.1860: Structural and Functional Features

NADP-Dependent Aldehyde Dehydrogenase from ArchaeonPyrobaculum sp.1860: Structural and Functional Features

A facile stable-isotope dilution method for determination of sphingosine phosphate lyase activity

Characterization of homologous sphingosine-1-phosphate lyase isoforms in the bacterial pathogen Burkholderia pseudomallei

Contact Info

Product

Resources

About