Posttranslational modification of serine to formylglycine in bacterial sulfatases. Recognition of the modification motif by the iron-sulfur protein AtsB.
Calpha-formylglycine is the catalytic residue of sulfatases. Formylglycine is generated by posttranslational modification of a cysteine (pro- and eukaryotes) or serine (prokaryotes) located in a conserved (C/S)XPXR motif. The modifying enzymes are unknown. AtsB, an iron-sulfur protein, is strictly required for modification of Ser(72) in the periplasmic sulfatase AtsA of Klebsiella pneumoniae. Here we show (i) that AtsB is a cytosolic protein acting on newly synthesized serine-type sulfatases, (ii) that AtsB-mediated FGly formation is dependent on AtsA's signal peptide, and (iii) that the cytosolic cysteine-type sulfatase of Pseudomonas aeruginosa can be converted into a substrate of AtsB if the cysteine is substituted by serine and a signal peptide is added. Thus, formylglycine formation in serine-type sulfatases depends both on AtsB and on the presence of a signal peptide, and AtsB can act on sulfatases of other species. AtsB physically interacts with AtsA in a Ser(72)-dependent manner, as shown in yeast two-hybrid and GST pull-down experiments. This strongly suggests that AtsB is the serine-modifying enzyme and that AtsB relies on a cytosolic function of the sulfatase's signal peptide.
Marquordt
Claudia
C
Institut für Biochemie und Molekulare Zellbiologie, Abt. Biochemie II, Universität Göttingen, Heinrich-Düker-Weg 12, Germany.
Fang
Qinghua
Q
Will
Elke
E
Peng
Jianhe
J
von Figura
Kurt
K
Dierks
Thomas
T
D000409:Alanine / Q000031:analogs & derivatives* / Q000737:chemistry; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001192:Arylsulfatases / Q000737:chemistry* / Q000378:metabolism; D015153:Blotting, Western; D003545:Cysteine / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D005998:Glycine / Q000031:analogs & derivatives* / Q000737:chemistry*; D007506:Iron-Sulfur Proteins / Q000737:chemistry* / Q000378:metabolism; D007711:Klebsiella pneumoniae / Q000378:metabolism; D008956:Models, Chemical; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals; D012694:Serine / Q000737:chemistry* / Q000378:metabolism; D013347:Subcellular Fractions; D013429:Sulfatases / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D001616:beta-Galactosidase / Q000378:metabolism
C471555:AtsA protein, Klebsiella pneumoniae; C471554:AtsB protein, Klebsiella pneumoniae; D007506:Iron-Sulfur Proteins; D010455:Peptides; D021382:Protein Sorting Signals; D012694:Serine; C524739:C(alpha)-formylglycine; D005982:Glutathione Transferase; D013429:Sulfatases; D001192:Arylsulfatases; D001616:beta-Galactosidase; D003545:Cysteine; D000409:Alanine; D005998:Glycine