The reaction catalyzed by Rv3406.

<p>The alpha carbon of an alkyl sulfate is oxidized by Rv3406 in the presence of α-ketoglutarate and spontaneously collapses to an aldehyde and sulfate, liberating CO₂ and succinate. The product formation was monitored using a coupled assay using LADH to reduce the aldehyde in a NADH dependent manner.</p

Biochemical characterization of Rv3406.

<p>(A) Rv3406 is an αKG and ascorbate dependent sulfatase. Black squares are the complete assay with Rv3406, 2-EHS, αKG, ascorbate and iron. Red triangles are without 2-EHS. Blue circles are without αKG. (B) Rv3406 is an iron dependent enzyme. (C) The rate of Rv3406 accelerates with the addition of ascorbate up to 1 mM. Rv3406 enzyme concentration was between 0.5 and 0.75 µM for all experiments.</p

Protein alignment of alkyl sulfatase enzymes with taurine dioxygenase enzymes.

<p><b>Enzymes in bold have been biochemically characterized.</b> (A) Alignment of disordered loop 1 where the red boxes are indicating the taurine binding residues in taurine dioxygenases and the analogous amino acids in alkyl sulfatase enzymes. (B) Alignment of disordered loop 2 where the red box is indicating the conserved phenylalanine in taurine dioxygenases and the analogous tyrosine in alkyl sulfate enzymes.</p

Rv3406 is essential in Mtb for growth on 2-EHS as the sole sulfur source.

<p>(A) Growth of Mtb strains using either 2-EHS alone or 2-EHS with sodium sulfate. (B) Growth of Mtb strains on <i>n</i>-heptyl sulfate or SDS. Data represents three biological replicates and error bars denote standard deviation. Asterisk indicates a p value of less than 0.005.</p

Conditional Glycosylation in Eukaryotic Cells Using a Biocompatible Chemical Inducer of Dimerization

Conditional Glycosylation in Eukaryotic Cells Using a Biocompatible Chemical Inducer of Dimerizatio

Sulfolipid-1 Biosynthesis Restricts <i>Mycobacterium tuberculosis</i> Growth in Human Macrophages

<i>Mycobacterium tuberculosis</i> (Mtb), the causative agent of tuberculosis, is a highly evolved human pathogen characterized by its formidable cell wall. Many unique lipids and glycolipids from the Mtb cell wall are thought to be virulence factors that mediate host–pathogen interactions. An intriguing example is Sulfolipid-1 (SL-1), a sulfated glycolipid that has been implicated in Mtb pathogenesis, although no direct role for SL-1 in virulence has been established. Previously, we described the biochemical activity of the sulfotransferase Stf0 that initiates SL-1 biosynthesis. Here we show that a <i>stf0</i>-deletion mutant exhibits augmented survival in human but not murine macrophages, suggesting that SL-1 negatively regulates the intracellular growth of Mtb in a species-specific manner. Furthermore, we demonstrate that SL-1 plays a role in mediating the susceptibility of Mtb to a human cationic antimicrobial peptide <i>in vitro</i>, despite being dispensable for maintaining overall cell envelope integrity. Thus, we hypothesize that the species-specific phenotype of the <i>stf0</i> mutant is reflective of differences in antimycobacterial effector mechanisms of macrophages