The pyruvate formate lyase family: sequences, structures and activation.
We cloned and expressed in Escherichia coli the Archaeglobus fulgidus gene that encodes pyruvate formate lyase 2 (PFL2). PFL2, despite its homology to the other glycyl radical enzymes, differs from them by exhibiting a completely different oligomerization. The most abundant form of PFL2 when expressed in E.coli is a trimer. The closest homologue of PFL2 with a known structure is E. coli PFL, which is a dimer. Sequence comparisons allowed us to reclassify PFL-like enzymes and the consensus sequences allowed us to propose an activation route for PFL-like glycyl radical enzymes. Surprisingly, most of the conserved residues in PFL-like enzymes appear to be involved in preserving the structure, rather than forming the active site.
Graduate School in Informational and Structural Biology, Institute of Biotechnology, University of Helsinki, PO Box 65, FIN-00014 Helsinki, Finland.
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Lehtiö
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Goldman
D000123:Acetyltransferases / Q000737:chemistry / Q000145:classification / Q000235:genetics*; D000595:Amino Acid Sequence; D019611:Archaeoglobus fulgidus / Q000201:enzymology* / Q000235:genetics; D020134:Catalytic Domain / Q000235:genetics; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D016384:Consensus Sequence / Q000235:genetics; D017124:Conserved Sequence / Q000235:genetics; D003545:Cysteine / Q000235:genetics; D019641:DNA, Archaeal / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics; D008027:Light; D008958:Models, Molecular; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D012542:Scattering, Radiation; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D019641:DNA, Archaeal; D021122:Protein Subunits; D011994:Recombinant Proteins; D000123:Acetyltransferases; C022560:formate C-acetyltransferase; D003545:Cysteine