The dimerization motif of cytosolic sulfotransferases.
Cytosolic sulfotransferases sulfate steroids such as estrogens and hydroxysteroids. The enzymes, including human estrogen sulfotransferase (hEST) and hydroxysteroid sulfotransferase (hHST), are generally homodimers in solution with mouse estrogen sulfotransferase (mEST) being one of few exceptions. To identify the amino acid residues responsible for the dimerization, eight residues on the surface of hEST were mutated to their counterparts in mEST and mutated hESTs were then analyzed by gel filtration chromatography. A single mutation of Val(269) to Glu was sufficient to convert hEST to a monomer and the corresponding mutation of Val(260) also altered hHST to a monomer. The hHST crystal structure revealed a short stretch of peptide with the side-chains from two hHST monomers forming a hydrophobic zipper-like structure enforced by ion pairs at both ends. This peptide consisted of 10 residues near the C-terminus that, including the critical Val residue, is conserved as KXXXTVXXXE in nearly all cytosolic sulfotransferases. When mEST underwent the double mutations Pro269Thr/Glu270Val dimerization resulted. Thus, the KXXXTVXXXE sequence appears to be the common protein-protein interaction motif that mediates the homo- as well as heterodimerization of cytosolic sulfotransferases.
Petrotchenko
E V
EV
Pharmacogenetics Section, Laboratory of Reproductive and Developmental Toxicology, NIEHS/NIH, Research Triangle Park, NC 27709, USA.
Pedersen
L C
LC
Borchers
C H
CH
Tomer
K B
KB
Negishi
M
M
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D002850:Chromatography, Gel; D003432:Cross-Linking Reagents / Q000378:metabolism; D018360:Crystallography, X-Ray; D003600:Cytosol / Q000201:enzymology*; D004252:DNA Mutational Analysis; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D005973:Glutamine / Q000737:chemistry; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D013058:Mass Spectrometry; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D012685:Sepharose / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D015238:Sulfotransferases / Q000737:chemistry* / Q000378:metabolism*; D014633:Valine / Q000737:chemistry
D003432:Cross-Linking Reagents; D005973:Glutamine; D012685:Sepharose; D015238:Sulfotransferases; D005978:Glutathione; D014633:Valine